EP4731223A1 - Morpholine and 1,4-oxazepane compounds and its use in therapy - Google Patents

Morpholine and 1,4-oxazepane compounds and its use in therapy

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Publication number
EP4731223A1
EP4731223A1 EP24736766.7A EP24736766A EP4731223A1 EP 4731223 A1 EP4731223 A1 EP 4731223A1 EP 24736766 A EP24736766 A EP 24736766A EP 4731223 A1 EP4731223 A1 EP 4731223A1
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Prior art keywords
methyl
piperidin
benzyl
oxazepane
oxazepan
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German (de)
French (fr)
Inventor
Cristian Gaspar GRIÑAN FERRE
Mercè PALLÀS LLIBERIA
Aina BELLVER SANCHIS
María Carmen ESCOLANO MIRÓN
Santiago VÁZQUEZ CRUZ
Carla BARBARACI
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Universitat de Barcelona UB
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Universitat de Barcelona UB
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Publication of EP4731223A1 publication Critical patent/EP4731223A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/553—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having at least one nitrogen and one oxygen as ring hetero atoms, e.g. loxapine, staurosporine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
    • A61P25/32—Alcohol-abuse
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid

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  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
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  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
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  • Hospice & Palliative Care (AREA)
  • Immunology (AREA)
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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Plural Heterocyclic Compounds (AREA)

Abstract

The present invention provides compounds of formula (I), or a salt, solvate or 5 stereoisomer thereof as G9a inhibitors, wherein: R1 is CRxR'xRy; A represents (CH2)m, wherein m is 0 or 1; Z represents (CH2)n, wherein n is 2 or 3; R2 is selected from the group consisting of: (a) an aromatic 5 or 6-membered ring system, wherein the members are selected from the group con sisting of: CRz1, S, N, NH, and O; and (b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRz2, S, N, NH, and O. The invention also provides uses of the compounds of formula (I) in the treatment of G9a-mediated diseases, as well as pharmaceutical compositions including these compounds. Advantageously, the compounds of the invention are highly potent and selective towards G9a target, among others.

Description

MORPHOLINE AND 1,4-OXAZEPANE COMPOUNDS AND ITS USE IN THERAPY
FIELD OF THE INVENTION
The present invention refers to new compounds which are potent and specific G9a inhibitors, as well as to pharmaceutical compositions comprising these compounds and uses thereof in the treatment or prevention of diseases or conditions related to G9a overexpression.
BACKGROUND
Histone methyltransferases (HMTs) catalyzes the methylation of specific lysines and arginines in histones H3 and H4. This histone methylation can result in either the activation or repression of gene transcription. Thus, dysregulation of methylation at specific histone sites (alterations in the “histone code”) has been implicated in many cancers. Hence, targeting HMT activity has been the subject of much investigation in the field of oncology.
One such HMT is Euchromatic histone methyltransferase 2 (EHMT2), also known as G9a. G9a is primarily responsible for the dimethylation of lysine 9 on histone H3 (H3K9).
G9a overexpression has already been reported in numerous cancers such as hematologic, breast, gastric, ovarian, prostate, lung, colorectal, liver, urinary bladder, and brain cancers has sparked interest in it for therapeutic development (Barghout S. H. et al., “Chemical biology and pharmacology of histone lysine methylation inhibitors”, BBA - Gene Regulatory Mechanisms, 2022, 1865, 194840); as well as involved in immune system activation (Scheer S. et al., “The Lysine Methyltransferase G9a in immune Cell Differentiation and Function”, Frontiers in Immunology, 2017, 8: 429); in memory function (K.K.L. et al., “Epigenetics and memory: Emerging role of histone lysine methyltransferase G9a/GLP complex as bidirectional regulator of synaptic plasticity”, Neurobiology of Learning and Memory, 2019, 159, 1-5); or Prader-Willi syndrome (Kim Y. et al., “Epigenetic therapy of Prader-Willi syndrome”, Translational Research, 2019, 106, 105-118).
While small molecule inhibitors of G9a have been reported as early as 2005, their effectiveness has largely been unsuccessful in G9a-mediated diseases. Accordingly, there is a need in the art for specific inhibitors of G9a.
SUMMARY OF THE INVENTION
The present inventors have developed new G9a inhibitors which solve the problems of the already known inhibitors.
In particular, it has been found that the compounds provided in the present invention are potent (Table 1) and highly selective (Table 2) G9a inhibitors.
Interestingly, the compounds of the invention were able to reduce H3K9me2 (H3K9me2 indicates dimethylation of lysine 9 on histone H3 protein subunit) levels, suggesting the G9a is the direct target.
The role of the compounds of the invention on G9a-related diseases was also investigated.
On the one hand, the effect on the impact A aggregates reduction was better for the compound of the invention in comparison with a well-established G9a inhibitor, UNC0638.
In addition, the compounds of the invention also exhibited a high PAMPA-BBB permeability compared to UNC0638 and no remarkable toxicity was found in the several tests performed (see for instance Table 5 below).
Remarkably, in vivo study in SAMP8, a well-established AD (Alzheimer's disease) mice model, showed better cognitive performance using NORT (Novel Object Recognition Task) and OLT (Object Location Task) as well as an enhancement in synaptic plasticity evaluated through dendritic spine length and density length that were increased in SAMP8 treated with the compound of the invention.
Altogether, the data provided herein support that the compound of the invention improves potency, selectivity, blood brain barrier permeability, good metabolism, and pharmacokinetic profile, rescuing the main AD hallmarks in different AD models. Thus, in a first aspect the present invention provides a compound of formula (I), or a salt, solvate or stereoisomer thereof: for use in therapy, wherein:
Ri is CRxR’xRy,
A represents (CH2)m, wherein m is 0 or 1 ,
Z represents (CH2)n, wherein n is 2 or 3,
R2 is selected from the group consisting of: a) an aromatic 5 or 6-membered ring system, wherein the members are selected from the group consisting of: CRzi, S, N, NH, and O; and b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRZ2, S, N, NH, and O;
Rx and R’x are the same or different, and are independently selected from -H, (C1- Cs)alkyl, (Ci-Cs)haloalkyl;
Ry is selected from -H, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, or an aromatic known ring system, which is selected from the group consisting of: a) an aromatic 6-membered ring system, wherein the members are selected from the group consisting of: CRZ3, and N; b) an aromatic 5-membered ring system, wherein the members are selected from the group consisting of: CRZ4, S, N, NH, and O; c) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings having 5 or 6 members selected from the group consisting of: CRzs, N and NH;
Rzi to RZ3 and Rzs are independently selected from the group consisting of: H; OH; CN; halogen; (Ci-Cw)alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more SU3; -0-(Ci-Cio)alkyl; -O-(Ci-Cw)haloalkyl; SF5; S(O)2Rs; NR4R5; CONReR?; and COORs; RZ4 is selected from the group consisting of: H; OH; CN; halogen; (Ci-Cw)haloalkyl; -O- (Ci-Cw)alkyl; -O-(Ci-Cw)haloalkyl; SF5; S(O)2R’3; NR^R’s; CONR’eR’?; and COOR’s;
R3 and R’3 are independently selected from (Ci-Cw)alkyl; (C3-C8)cycloalkyl; aryl; or heteroaryl;
R4, Rs, Re, R7, R’4, R’s, R’e and R’7, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl;
Rs and R’s are independently selected from the group consisting of: selected from the group consisting of -H, (Ci-Cw)alkyl; (C3-Cs)cycloalkyl; and (Ci-Cw)haloalkyl;
“aryl” means an aromatic ring system comprising 6 CRc members, being Rc selected from H, halogen, CN, nitro, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, -O-(Ci-Cs)alkyl, or -O-(Ci- Cs)haloalkyl;
“heteroaryl” means an aromatic ring system comprising 5 or 6 members selected from the group consisting of: CRd, O, N, NH, and S; being Rd selected from H, halogen, CN, nitro, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, -O-(Ci-Cs)alkyl, -O-(Ci-Cs)haloalkyl;
Sui to Su3 are independently selected from the group consisting of: halogen, CN, nitro, (Ci-Ce)haloalkyl, (Ci-Ce)alkyl, (Ci-Ce)hydroxyalkyl, (Ci-Ce)alkoxy, (Ci-Ce)alkylsulfinyl, (Ci-Ce)alkylsulfonyl, (Ci-Ce)alkylcarbonyl, (Ci-Ce)alkoxycarbonyl, carbamoyl, /\/-(Ci- C4)alkylcarbamoyl, /V,/V-di-(Ci-C4)alkylcarbamoyl, (Ci-Ce)alkylcarbonyloxy, (C3- C6)cycloalkyl, phenyl, benzyl, phenoxy, benzyloxy, anilino, /V-methylanilino, phenylmercapto, phenylsulfonyl, phenylsulfinyl, sulfamoyl, /V-(Ci-C4)alkylsulfamoyl, and /V,/V-di-(Ci-C4)alkylsulfamoyl.
In a second aspect the present invention provides the compound of formula (I) as defined in the first aspect of the invention, for use in preventing or treating a G9a-mediated disorder in a subject in need thereof. This aspect can alternatively be formulated as the use of a compound of formula (I) as defined in the first aspect of the invention for the manufacture of a medicament for the prevention or treatment of a G9a-mediated disorder. This aspect can also alternatively be formulated as a method for the prevention or treatment of a G9a-mediated disorder, the method comprising administering a therapeutically effective amount of the compound of formula (I) as defined in the first aspect of the invention, to a subject in need thereof. In a third aspect the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in the first aspect of the invention, and one or more pharmaceutically acceptable excipients or carriers.
In a fourth aspect the present invention provides a compound of formula (Ibis), a salt, solvate or stereoisomer thereof: wherein Ri, A, and Z are as defined in the first aspect of the invention,
R2bis is selected from the group consisting of: a) an aromatic 6-membered ring system, wherein the members are selected from the group consisting of: CRzi, and N; and b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRZ2, S, N, NH, and O; and
CRzi and CRZ2 are as defined above; provided that R2bis is other than 3-hydroxyphenyl; or alternatively provided that when Ri is benzyl, m is 1 and n is 3, R2bis is other than 3-hydroxyphenyl; or alternatively provided that the compound of formula (Ibis) is other than 3-{1-[(4-benzyl-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}phenol.
In a final aspect the present invention provides a process for preparing the compounds of formula (I) of the invention, by reacting a compound of formula (II) with one of formula (HI): wherein “X”, represents a leaving group such as halogen or sulfonate (e.g. such as mesylate, tosylate, triflate or the like), particularly X represents Cl; and A, Z, Ri and R2 are as defined in the first aspect of the invention.
DETAILED DESCRIPTION OF THE INVENTION
Terms not specifically defined herein should be given the meanings that would be given to them by one of skill in the art in light of the disclosure and the context. As used in the specification, however, unless specified to the contrary, the following terms have the meaning indicated and the following conventions are adhered to.
Throughout the present specification and the accompanying clauses, the words "comprise" and variations such as "comprises", "comprising" are to be interpreted inclusively. That is, these words are intended to convey the possible inclusion of other elements or integers not specifically recited, where the context allows. The word “comprise” also includes the term “consists of”.
For the purposes of the present invention, any ranges given include both the lower and the upper end-points of the range.
In a first aspect the present invention provides the use of compounds of formula (I) in therapy.
In the context of the invention, the term "alkyl" refers to a straight or branched hydrocarbon chain radical containing no unsaturation, and which is attached to the rest of the molecule by a single bond. Typical alkyl groups have from 1 to about 10, 1 to about 8, or 1 to about 6 carbon atoms, e. g., methyl, ethyl, n-propyl, /-propyl, n-butyl, f-butyl, n- pentyl, etc. If substituted by cycloalkyl, it corresponds to a "cycloalkylalkyl" radical, such as cyclopropyl methyl. If substituted by aryl, it corresponds to an "arylalkyl" radical, such as benzyl, benzhydryl or phenethyl. If substituted by heterocyclyl, it corresponds to a "heterocyclylalkyl" radical.
In the context of the invention, the term "alkenyl" refers to a straight or branched hydrocarbon chain radical containing at least two carbon atoms and at least one C=C double bond, and which is attached to the rest of the molecule by a single bond. Typical alkenyl radicals have from 2 to about 10, 2 to about 8 or 2 to about 6 carbon atoms. In a particular embodiment, the alkenyl group is vinyl, 1-methyl-ethenyl, 1 -propenyl, 2- propenyl, or butenyl.
In the context of the invention, the term "alkynyl" refers to a straight or branched hydrocarbon chain radical containing one or more C=C triple bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, 2-propynyl, and 2-butynyl.
In the context of the invention, the term “hydroxyalkyl” refers to a straight or branched hydrocarbon chain radical containing no unsaturation, wherein one or more of the hydrogens are replaced by -OH. Illustrative non-limitative examples of hydroxyalkyl are methanol, ethanol, isopropanol, isobutanol or f-butanol, among others.
In the context of the invention, the term “alkoxy” refers to a -O-alkyl, wherein “alkyl” is as defined above. Illustrative non-limitative examples of hydroxyalkyl are methoxy, ethoxy, or f-butoxy, among others.
In the context of the invention, the term “alkylsulfinyl” refers alkyl-SO-, being “alkyl” as defined above.
In the context of the invention, the term “alkylsulfonyl” refers to alkyl-SCh-, being “alkyl” as defined above.
In the context of the invention, the term “alkylcarbonyl” means a straight or branched hydrocarbon chain radical containing no unsaturation, where the alkyl chain is interrupted with a carbonyl (C=O) group (i.e., an alkyl-C(O)-alkylene-group). Representative alkylcarbonyl groups include methylcarbonymethyl, ethylcarbonylmethyl, methylcarbonylethyl, (2-methylpropyl)carbonylmethyl, and the like.
In the context of the invention, the term “alkoxycarbonyl” refers to an “alkoxy”, as defined above, where the alkyl chain is interrupted with a carbonyl (C=O) group.
In the context of the invention, the term “carbamoyl” refers to -C(O)NH2.
In the context of the invention, the term “alkylcarbonyloxy” refers to -C(O)-O-alkyl, being alkyl as defined above.
In the context of the invention, the term “sulfamoyl” refers to -S(O)2NH2.
In the context of the invention, the term "halogen" refers to bromo, chloro, iodo or fluoro. In the context of the invention, the term “haloalkyl” refers to a straight or branched hydrocarbon chain radical containing no unsaturation, wherein one or more of the hydrogen atoms are replaced by halogen. Illustrative non-limitative examples of haloalkyl are chloromethyl, trifluoromethyl, 1-chloro-2-fluoroethyl, and the like.
In the context of the present invention, the term "salt" must be understood as any form of a compound used in accordance with this invention in which said compound is in ionic form or is charged and coupled to a counter-ion (a cation or anion) or is in solution. This definition also includes quaternary ammonium salts and complexes of the active molecule with other molecules and ions, particularly, complexes formed via ionic interactions. The definition includes in particular physiologically acceptable salts; this term must be understood as equivalent to "pharmacologically acceptable salts" or "pharmaceutically acceptable salts".
In the context of the present invention, the term "pharmaceutically acceptable salts" means any salt that is tolerated physiologically (normally meaning that it is not toxic, particularly, as a result of the counter-ion) when used in an appropriate manner for a treatment, applied or used, particularly, in humans and/or mammals. These physiologically acceptable salts may be formed with cations or bases and, in the context of this invention, are understood to be salts formed by at least one compound used in accordance with the invention -normally an acid (deprotonated)- such as an anion and at least one physiologically tolerated cation, preferably inorganic, particularly when used in humans and/or mammals. Salts with alkali and alkali earth metals are preferred particularly, as well as those formed with ammonium cations (NH4 +). Preferred salts are those formed with (mono) or (di)sodium, (mono) or (di)potassium, magnesium or calcium. These physiologically acceptable salts may also be formed with anions or acids and, in the context of this invention, are understood as being salts formed by at least one compound used in accordance with the invention - normally protonated, for example in nitrogen - such as a cation and at least one physiologically tolerated anion, particularly when used on humans and/or mammals. This definition specifically includes in the context of this invention a salt formed by a physiologically tolerated acid, i.e., salts of a specific active compound with physiologically tolerated organic or inorganic acids - particularly when used on humans and/or mammals. Examples of this type of salts are those formed with: hydrochloric acid, hydrobromic acid, sulphuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid or citric acid. In the context of the present invention, the term "solvate" should be understood as meaning any form a compound in accordance with the invention in which said compound is bonded by a non-covalent bond to another molecule (normally a polar solvent), including especially hydrates and alcoholates, like for example, methanolate. A preferred solvate is the hydrate.
The term "prodrug" is used in its broadest sense and encompasses those derivatives that are converted in vivo to the compounds of the invention. Examples of prodrugs include, but are not limited to, derivatives and metabolites of the compounds of formula (I) that include biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogues. Preferably, prodrugs of compounds with carboxyl functional groups are the lower alkyl esters of the carboxylic acid. The carboxylate esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule. Prodrugs can typically be prepared using well-known methods.
Any compound of formula (I) referred to herein is intended to represent such specific compound as well as certain variations or forms. In particular, compounds referred to herein may have asymmetric centres and therefore exist in different enantiomeric or diastereomeric forms. Thus, any given compound of formula (I) referred to herein is intended to represent any one of a racemate, one or more enantiomeric forms, one or more diastereomeric forms, and mixtures thereof. Likewise, stereoisomerism or geometric isomerism about the double bond is also possible, therefore in some cases the molecule could exist as (E)-isomer or (Z)-isomer (trans and cis isomers). If the molecule contains several double bonds, each double bond will have its own stereoisomerism, that could be the same as, or different to, the stereoisomerism of the other double bonds of the molecule. Furthermore, compounds referred to herein may exist as atropisomers. All the stereoisomers including enantiomers, diastereoisomers, geometric isomers and atropisomers of the compounds referred to herein, and mixtures thereof, are considered within the scope of the present invention.
Furthermore, any compound of formula (I) referred to herein may exist as tautomer. Specifically, the term tautomer refers to one of two or more structural isomers of a compound that exist in equilibrium and are readily converted from one isomeric form to another. In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein m is 1. In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein n is 3. In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein m is 1 and n is 3. In another embodiment of the first aspect of the invention, the compound of formula (I) is one wherein m is 1 and n is 2. In another embodiment of the first aspect of the invention, the compound of formula (I) is one wherein m is 0 and n is 3.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein Rx and R’x are the same. Particularly, Rx and R’x are H.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein Ry is an aromatic 6-membered ring system.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein the aromatic ring consists of 6 members, which are the same or different, and are CRZ3.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein Ry represents phenyl.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein R2 is an aromatic 5 or 6-membered ring system. Particularly, R2 is an aromatic 6-membered ring system.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one wherein Rzi is selected from H; OH; halogen; (Ci-Cio)alkyl; (C2-Cio)alkenyl; (C2- Cw)alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more SU3; -0-(Ci-Cio)alkyl; -0-(Ci-Cio)haloalkyl; or NR4R5. Particularly, Rzi is selected from H; OH; halogen; (Ci- Cio)alkyl; -0-(Ci-Cio)alkyl; or NR4R5.
In one embodiment of the first aspect of the invention, the compound of formula (I) is one selected from the group consisting of:
1. 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1 ,4-oxazepane,
2. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 3. 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4. 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
5. 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
6. 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
7. 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
8. 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
9. 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
10. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
11. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
12. 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
13. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
14. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
15. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline;
16. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile,
17. 2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
18. 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
19. 3-{ 1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline,
20. 4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
21 . 4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1 -yl]methyl}- 1 ,4-oxazepane,
22. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
23. 4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
24. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
25. methyl 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate,
26. 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1-yl)methyl}-1 ,4-oxazepane,
27. 2-{[4-(1/7-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
28. 4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
29. 2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
30. 3-{1-[(4-(4-chlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
31. 3-{1-[(4-(4-methylbenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
32. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine,
33. 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
34. 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine,
35. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine,
36. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine,
37. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
38. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-methylphenol, 39. 5-{1 -[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-fluorophenol,
40. 3-{1-[(4-(4-methoxybenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
41. 3-{1-[(4-(3,4-dichlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
42. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
43. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
44. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
45. 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
46. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
47. 4-methyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
48. 2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
49. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol,
50. 4-{1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, and any salt, solvate or stereoisomer thereof
G9a and GLP (G9a like protein) are major enzymes that catalyze the mono- and dimethylation of a lysine residue at position 9 of histone H3 (H3K9me1 and H3K9me2). These enzymes are also known as EHMT2 and EHMT1 (euchromatin histone-lysine N- methyltransferases 2 and 1).
H3K9me2 is an epigenetic mark related to transcriptional repression. G9a and GLP are involved in epigenetic transcriptional repression through H3K9me2.
The inhibition of G9a has been widely reported as useful for the control of biological processes, such as cell proliferation and cell differentiation, mediated by transcriptional repression by H3K9me2.
The present invention provides compounds which are potent and selective G9a inhibitors. Not only that, but the inventors have also confirmed that this is mainly due to the effect on H3K9me2.
As it is shown below the compounds of the invention inhibit the dimethylation of lysine 9 on histone H3, by inhibiting G9a, which catalyzes the mono-, di-, and/or trimethylation of lysine. As used herein, the term “inhibit” refers to a reduction or decrease in a quality or quantity, compared to a baseline. In one embodiment, the G9a-mediated disorder is associated with overexpression of G9a. G9a overexpression has already been reported in numerous cancers such as hematologic, breast, gastric, ovarian, prostate, lung, colorectal, liver, urinary bladder, and brain cancers has sparked interest in it for therapeutic development (Barghout S. H. et al. , “Chemical biology and pharmacology of histone lysine methylation inhibitors”, BBA - Gene Regulatory Mechanisms, 2022, 1865, 194840); as well as involved in immune system activation (Scheer S. et al., “The Lysine Methyltransferase G9a in immune Cell Differentiation and Function”, Frontiers in Immunology, 2017, 8: 429); in memory function (K.K.L. et al., “Epigenetics and memory: Emerging role of histone lysine methyltransferase G9a/GLP complex as bidirectional regulator of synaptic plasticity”, Neurobiology of Learning and Memory, 2019, 159, 1-5); or Prader-Willi syndrome (Kim Y. et al., “Epigenetic therapy of Prader-Willi syndrome”, Translational Research, 2019, 106, 105-118).
The compound of formula (I), in the context of the invention, can be administered in combination with any other therapeutic agent for preventing or treating the disorder, including G9a inhibitors, where the combination of the drugs together are safer or more effective than either drug alone. Such other drug(s) may be administered, by a route and in an amount commonly used therefore, contemporaneously or sequentially with the composition. When a compound of Formula I is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of formula I is preferred. However, the combination therapy may also include therapies in which the compound of Formula I and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the present invention and the other active ingredients may be used in lower doses than when each is used singly. Accordingly, the pharmaceutical compositions of the present invention include those that contain one or more other active ingredients, in addition to a compound of Formula I. The above combinations include combinations of a compound of the present invention not only with one other active compound, but also with two or more other active compounds. For example, the compound of Formula I can be combined with a variety of different anti-cancer drugs such as chemotherapeutics, anti-tumor agents, and anti-proliferative agents.
Further, the compound of Formula I can be combined with the following, but not limited to, actinomycins, alkylating agents, anthracyclines, antifolates, antiestrogen agents, antimetabolites, anti-androgens, antimicrotubule agents, aromatase inhibitors, bleomycins, Ca2+ adenosine triphosphate (ATP)ase inhibitors, cytosine analogs, deltoids/retinoids, dihydrofolate reductase inhibitors, deoxyribonucleic acid (DNA) topoisomerase inhibitors, dopaminergic neurotoxins, glucocorticoids, histone deacetylase inhibitors, hormonal therapies, immunotherapeutic agents, inosine monophosphate (IMP) dehydrogenase inhibitors, isoprenylation inhibitors, luteinizing hormone-releasing hormone agonists, mammalian target of rapamycin (mtor) inhibitors, multi-drug resistance (MDR) inhibitors, mitomycins, photodynamic therapies, proteasome inhibitors, platinum containing compounds, radiation, receptor tyrosine kinase inhibitors, ribonucleotide reductase inhibitors, thrombospondin mimetics, uracil analogs, vinca alkaloids, and vitamin D3 analogs such as, but not limited to, y-radiation or an additional chemotherapeutic agent or additional chemotherapeutic agents such as N-Ac-Sar-Gly- Val-D-allolle-Thr-Nva-lle-Arg-Pro-NHCH2CH3 or a salt thereof, actinomycin D, AG13736, alisertib, 17-allylamino-17-demethoxygeldanamycin, 9-aminocamptothecin, /\/-(4-(3- amino-1/7-indazol-4-yl)phenyl}-/\/'-(2-fluoro-5-methylphenyl)urea or a salt thereof, /\/-(4- (4-aminothieno[2,3-c(]pyrimidin-5-yl)phenyl}-/\/'-(2-fluoro-5-(trifluoromethyl)phenyl)urea or a salt thereof, anastozole, AP-23573, asparaginase, azacitidine, bevacizurnab, bicalutamide, bleomycin a2, bleomycin b2, bortezemib, busulfan, campathecins, carboplatin, carmustine (BCNll), CB1093, cetuximab, CHOP (C: Cytoxan® (cyclophosphamide); H: Adriamycin® (hydroxydoxorubicin); O: Vincristine (Oncovin®); P: prednisone), chlorambucil, CHIR258, cisplatin, CNF-101 , CNF-1001 , CNF-2024, CP547632, crisnatol, cytarabine, cyclophosphamide, cytosine arabinoside, daunorubicin, dacarbazine, dactinomycin, dasatinib, daunorubicin, deferoxamine, demethoxyhypocrellin A, depsipeptide, dexamethasone, 17-dimethylaminoethylamino- 17-demethoxygeldanamycin, docetaxel, doxifluridine, doxorubicin, EB 1089, epothilone D, epirubicin, 5-ethynyl-1-13-D-ribofuranosylimidazole-4-carboxamide (EICAR), IS erlotinib, etoposide, everolimus, 5-fluorouracil (5-Fll), floxuridine, fludarabine, flutamide, gefitinib, geldanamycin, gemcitabine, goserelin, /\/-(2-(4-hydroxyanilino}-3-pyridinyl}-4- methoxybenzenesulfonamide or a salt thereof, hydroxyurea, idarubicin, ifosfamide, imatinab, interferon-a, interferon-y, I PI-504, irinotecan, KH 1060, lapatanib, leucovorin calcium, LAQ824, leuprolide acetate, letrozole, lomustine (CCNll), lovastatin, megestrol, melphalan, mercaptopurine, methotrexate, 1-methyl-4-phyenylpyridinium, MG132, mitomycin, mitoxantrone, MLN4924, MLN518, MS-275, mycophenolic acid, mitomycin C, nitrosoureas, oprelvekin, oxaliplatin, paclitaxel, PD98059, peplomycin, photosensitizer Pc4, phtalocyanine, pirarubicin, plicamycin, prednisone, procarbizine, PTK787, PU24FC1 , PU3, radicicol, raloxifene, rapamycin, ratitrexed, retinoids such as pheuretinide, ribavirin, rituximab (Rituxin®), sorafenib, staurosporine, steroids such as dexamethasone and prednisone, suberoylanilide hydroxamic acid, sunitinib, tamoxifen, taxol, temozolamide, temsirolimus, teniposide, thapsigargin, thioguanine, thrombospondin-1 , tiazofurin, topotecan, trapoxin, trastuzumab, treosulfan, trichostatin A, trimetrexate, trofosfamide, tumor necrosis factor, valproic acid, vemurafenib, VER49009, verapamil, vertoporfin, vinblastine, vincristine, vindesine, vinorelbine vitamin D3, VX-680, zactima, ZK-EPO, zorubicin or combinations thereof.
Likewise, compounds of the present invention may be used in combination with other drugs that are used in the prevention, treatment, control, amelioration, or reduction of risk of the diseases or conditions for which compounds of the present invention are useful as described above. Such other drugs may be administered, by a route and in an amount commonly used therefore, contemporaneously or sequentially with a compound of the present invention. When a compound of the present invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound of the present invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a compound of the present invention.
The weight ratio of the compound of the compound of the present invention to the second active ingredient may be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when a compound of the present invention is combined with another agent, the weight ratio of the compound of the present invention to the other agent will generally range from about 1000:1 to about 1 :1000, preferably about 200:1 to about 1 :200. Combinations of a compound of the present invention and other active ingredients will generally also be within the aforementioned range, but in each case, an effective dose of each active ingredient should be used. In such combinations the compound of the present invention and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).
The skilled person will routinely adjust the amounts and appropriate route of administration according to their expertise and the particular situation of the subject. In one embodiment, the disease is selected from the group consisting of Alzheimer’s disease (AD), amyotrophic lateral sclerosis, age-related tau astrogliopathy, aortic amyloidosis, argyrophilic grain disease, British familial dementia, cardiac amyloidosis, cerebral amyloid angiopathy, chronic traumatic encephalopathy, corneal dystrophies, corticobasal degeneration, Creutzfeldt-Jakob disease, Danish familial dementia, Down syndrome, familial amyloidosis, familial corneal amyloidosis, fatal insomnia, frontotemporal dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, hereditary cerebral hemorrhage with amyloidosis, Huntington’s disease, inflammation-associated amyloidosis, kuru, Lewy bodies dementia, Mediterranean fever, Niemann-Pick disease type C, Parkinson’s disease, Pick’s disease, primary age-related tauopathy, progressive subcortical gliosis, progressive supranuclear palsy systemic amyloidosis, including transthyretin-associated amyloidosis and light-chain amyloidosis, subacute sclerosing panencephalitis and tuberous sclerosis.
In another embodiment, the disorder is a blood disorder, such as sickle cell anemia or b- thalassemia or hematological cancer.
In another embodiment the disorder is a cancer, such as lymphoma, leukemia, melanoma, breast cancer, ovarian cancer, hepatocellular carcinoma, prostate carcinoma, lung cancer, brain cancer, or hematological cancer.
In another embodiment the disorder is an immune-mediated disease.
In another embodiment the compounds are used for reducing substance (alcohol) consumption by a subject.
In another embodiment the disorder is an imprinting disorder, such as Prader-Willi syndrome, transient neonatal diabetes mellitus, Silver-Russell syndrome, Albright hereditary osteodystrophy, pseudohypoparathyroidism, Birk-Barel mental retardation, Beckwith-Wiedemann syndrome, Temple syndrome, Kagami-Ogata syndrome, Angelman syndrome, precocious puberty, Schaaf-Yang syndrome, sporadic pseudohypoparathyroidism lb, or maternal uniparental disomy of chromosome 20 syndrome.
In a further aspect the invention provides a process for preparing the compounds of formula (I), by reacting a compound of formula (II) with one of formula (III). The reaction can take place in the presence of a base, such as KHCO3, NaHCCh, or the like, in a polar solvent such as MeCN, DMF, DMA, dichloromethane and, optionally, Nal as a catalyst, at a temperature around 70-90°C. Particular embodiments are provided in the examples below.
The compounds may be isolated and purified by methods well-known to those skilled in the art of organic synthesis. Examples of conventional methods for isolating and purifying compounds can include, but are not limited to, chromatography on solid supports such as silica gel, alumina, or silica derivatized with alkylsilane groups, by recrystallization at high or low temperature with an optional pretreatment with activated carbon, thin-layer chromatography, distillation at various pressures, sublimation under vacuum, and trituration.
Optimum reaction conditions and reaction times for each individual step can vary depending on the particular reactants employed and substituents present in the reactants used. Unless otherwise specified, solvents, temperatures and other reaction conditions can be readily selected by one of ordinary skill in the art. Specific procedures are provided in the Examples section. Reactions can be worked up in the conventional manner, e.g. by eliminating the solvent from the residue and further purified according to methodologies generally known in the art such as, but not limited to, crystallization, distillation, extraction, trituration and chromatography. Unless otherwise described, the starting materials and reagents are either commercially available or can be prepared by one skilled in the art from commercially available materials using methods described in the chemical literature.
Routine experimentations, including appropriate manipulation of the reaction conditions, reagents and sequence of the synthetic route, protection of any chemical functionality that cannot be compatible with the reaction conditions, and deprotection at a suitable point in the reaction sequence of the method are included in the scope of the invention. Suitable protecting groups and the methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art. Synthesis of the compounds of the invention can be accomplished by methods analogous to the one described in the synthetic scheme hereinabove and in specific examples provided below.
Starting materials, if not commercially available, can be prepared by procedures selected from standard organic chemical techniques, techniques that are analogous to the synthesis of known, structurally similar compounds, or techniques that are analogous to the above described schemes or the procedures described in the synthetic examples section.
When an optically active form of a compound of the invention is required, it can be obtained by carrying out one of the procedures described herein using an optically active starting material (prepared, for example, by asymmetric induction of a suitable reaction step), or by resolution of a mixture of the stereoisomers of the compound or intermediates using a standard procedure (such as chromatographic separation, recrystallization or enzymatic resolution).
Similarly, when a pure geometric isomer of a compound of the invention is required, it can be obtained by carrying out one of the above procedures using a pure geometric isomer as a starting material, or by resolution of a mixture of the geometric isomers of the compound or intermediates using a standard procedure such as chromatographic separation.
In a further aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compounds of the invention (I) as well as solvates, salts or stereoisomers thereof, as defined above. All the embodiments provided above, under the first aspect of the invention, concerning the compounds of formula (I), are also embodiments of the pharmaceutical composition of the invention.
By “therapeutically effective amount”, it is understood the amount of the compound(s) that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disease which is addressed.
The precise therapeutic dose of the component(s), as well as the amount of the compound(s) of the invention, may depend on several variables. Some of these would be: route of administration, time of drug release (e.g., instant or extended), administration schedule, pain severity, condition of the patient, and the like.
The pharmaceutical compositions can be prepared as a liquid, semi-solid or solid dosage form, for example in the form of solutions for injection, drops, juices, syrups, sprays, suspensions, tablets, patches, capsules, dressings, suppositories, ointments, creams, lotions, gels, emulsions, aerosols or in multiparticulate form, for example in the form of pills or granules, if appropriate compressed into tablets, decanted into capsules or suspended in a liquid, or administered as such.
These compositions can be prepared with the aid of conventional means, devices, methods or processes known in the art. Pharmaceutically acceptable adjuvants, vehicles or excipients which may be used in such compositions are adjuvants, vehicles or excipients known to those skilled in the art or commonly used in the preparation of therapeutic compositions, which may be selected, for example, from the group consisting of excipients, fillers, solvents, diluents, surfactants, colorants, preservatives, disintegrants, sliding agents, lubricants, flavoring agents or binders.
The term "pharmaceutically acceptable" refers to pharmaceutically acceptable materials, compositions or vehicles. Each component must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the pharmaceutical composition. It must also be suitable for use in contact with the tissue or organ of humans without excessive toxicity, irritation, allergic response, immunogenicity or other problems or complications commensurate with a reasonable benefit/risk ratio in animals and, particularly, in humans.
As used herein, the term “animal” or “subject” or “patient” shall refer to a vertebrate animal. Such animals include both domestic animals; for example, livestock, laboratory animals and household pets, and non-domestic animals such as wildlife. In one embodiment, the animal is a vertebrate. In a particular embodiment the animal is a domestic mammal or a human.
The selection of physiologically compatible adjuvants or the number of adjuvants to be used depends on the form of administration of the pharmaceutical composition, i.e., oral, subcutaneous, parenteral, intravenous, intraperitoneal, intradermal, intramuscular, intranasal, buccal, rectal, otic or intratympanic. Preparations in the form of tablets, dragees, capsules, granules, pills, drops, in particular otic drops, juices or syrups are preferably suitable for oral administration; solutions, suspensions, easily reconstitutable dry preparations or also sprays are preferably suitable for parenteral, topical or inhalation administration. The compounds in accordance with the invention used in the pharmaceutical composition in accordance with the invention in a depot, in a dissolved form or in a dressing, or if appropriate having added other agents favoring penetration into the skin, are preparations suitable for percutaneous administration. The preparation forms administrable orally or percutaneously can also release the respective compound according to the invention in a delayed form.
For instance, for oral administration in the form of a tablet or capsule, the active drug components can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulphate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and colouring agents can also be incorporated into the mixture. Suitable binders include starch, gelatine, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
Gelatine capsules contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.
Liquid dosage forms for oral administration can contain colouring and flavouring to increase patient acceptance.
The dosage administered of the pharmaceutical composition will, of course, vary depending on the use and known factors such as the age, health, and weight of the recipient; nature and extent of symptoms, concurrent treatments, if any, frequency of treatment, and the effect desired. The recipient may be any type of mammal, but is preferably a human.
In a fourth aspect, the present invention provides a compound of formula (Ibis). All the embodiments, provided under the first aspect of the invention, regarding A, Z, and Ri, are also embodiments of the compound of formula (Ibis) of the fourth aspect of the invention.
In one embodiment of the fourth aspect, R2bis is an aromatic 6-membered ring system.
In another embodiment of the fourth aspect, R2bis is an aromatic 6-membered ring system wherein all six members are the same or different and represent CRzi, being Rzi as defined in the first aspect of the invention or any of the embodiments provided above. In one embodiment of the fourth aspect, m is 1. In another embodiment of the fourth aspect, n is 3. In another embodiment of the fourth aspect, m is 1 and n is 3.
In one embodiment of the fourth aspect, Rx and R’x are the same. Particularly, Rx and R’x are H.
In one embodiment of the fourth aspect, Ry is an aromatic 6-membered ring system. Particularly, Ry is an aromatic ring consisting of 6 members, which are the same or different, and are CRZ3; particularly, they are CH.
In one embodiment of the fourth aspect, Rzi is selected from H; OH; halogen; (Ci- Cw)alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more Sus; -0-(Ci-Cio)alkyl; -O-(Ci-Cw)haloalkyl; or NR4R5. Particularly, Rzi is selected from H; OH; halogen; (Ci-Cw)alkyl; -0-(Ci-Cio)alkyl; or NR4R5.
In another embodiment of the fourth aspect, the compound is selected from:
1. 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1 ,4-oxazepane,
2. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
3. 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4. 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
5. 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
6. 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
7. 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
8. 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
9. 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
10. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
11 . 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
12. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
13. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
14. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline;
15. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile,
16. 2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
17. 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol, 18. 3-{ 1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline,
19. 4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
20. 4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
21. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
22. 4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
23. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
24. methyl 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate,
25. 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1-yl)methyl}-1 ,4-oxazepane,
26. 2-{[4-(1/7-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
27. 4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
28. 2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
29. 3-{1-[(4-(4-chlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
30. 3-{1-[(4-(4-methylbenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
31. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine,
32. 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
33. 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine,
34. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine,
35. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine,
36. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
37. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-methylphenol,
38. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-fluorophenol,
39. 3-{1-[(4-(4-methoxybenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
40. 3-{1-[(4-(3,4-dichlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
41. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
42. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
43. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
44. 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
45. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
46. 4-methyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
47. 2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
48. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol,
49. 4-{1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, or any salt, solvate or stereoisomer thereof. To those skilled in the art, other objects, advantages or features of the invention will be apparent in part from the description or in part from the practice of the invention. The following examples are provided by way of illustration or are not intended to be limiting of the present invention.
EXAMPLES
General procedure:
To a solution of 4-substituted piperidine (1 mmol) in MeCN, KHCO3 (3 mmol) and Nal (catalytic amount) were added. The solution was warmed up at 40°C and, after a few minutes, a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (1.31 mmol) in MeCN was added dropwise. The temperature was raised to 80°C and the reaction mixture was stirred overnight at 80°C. Then, a saturated solution of NaHCCh and EtOAc were added, and the phases were separated. The organic phase was washed with a saturated solution of NaHCOs, dried over anh. Na2SC>4, filtered and evaporated. The crude mixture was purified by flash column chromatography (CombiFlash®) using as eluent system DCM/MeOH or EtOAc/MeOH, furnishing the desired compounds as coloured oils.
Synthesis of 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1,4-oxazepane, compound 1.
From 4-phenylpiperidine (52 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1 ,4-oxazepane (77 mg, 66% yield) was obtained after chromatography DCM/MeOH (96:4). IR (ATR): 3026, 2929, 2849, 2809, 1602, 1493, 1452, 1353, 1259, 1137, 1091 , 1066, 1028, 993, 738, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H33N2O+H]+: 365.2587, found: 365.2586.
Synthesis of 4-benzyl-2-{[4-(4-chlorophenyl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 2.
From 4-(4-chlorophenyl)piperidine (63 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}- 1 ,4-oxazepane (89 mg, 70% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 3026, 2929, 2849, 2809, 1602, 1493, 1452, 1353, 1259, 1137, 1091 , 1066, 1028, 993, 738, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32CIN2O+H]+: 399.2198, found: 399.2196.
Synthesis of 4-benzyl-2-{[4-(4-bromophenyl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 3.
From 4-(4-bromophenyl)piperidine (77 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (125 mg, 1.25 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1- yl]methyl}-1 , 4-oxazepane (100 mg, 71 % yield) was obtained after column chromatography DCM/MeOH (95:5). IR (ATR): 3025, 2932, 2808, 1647, 1489, 1451 , 1352, 1137, 1089, 1073, 1008, 819, 736, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32BrN2O+H]+: 443.1693, found: 443.1690.
Synthesis of 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1, 4-oxazepane, compound 4.
From 4-(4-fluorophenyl)piperidine (54 pL, 0.32 mmol) in MeCN (4 mL), KHCO3 (125 mg, 1.25 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}- 1 , 4-oxazepane (59 mg, 48% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 3038, 2933, 2813, 1603, 1510, 1441 , 1352, 1250, 1216, 1162, 1097, 837, 738, 699 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32FN2O+H]+: 383.2493, found: 383.2502.
Synthesis of 4-benzyl-2-{[4-(3-bromophenyl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 5.
From 4-(3-bromophenyl)piperidine (77 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL) 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}- 1 , 4-oxazepane (97 mg, 68% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 2933, 2808, 1565, 1451 , 1353, 1137, 1088, 1071 , 995, 779, 743, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32BrN2O+H]+: 443.1693, found: 443.1693.
Synthesis of 4-benzyl-2-{[4-(3, 5-dichlorophenyl)piperidin-1-yl]methyl}- 1, 4-oxazepane, compound 6.
From 4-(3,5-dichlorophenyl)piperidine (74 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1- yl]methyl}-1 ,4-oxazepane (101 mg, 73% yield) as obtained after chromatography DCM/MeOH (97:3). IR (ATR): 2937, 2793, 1564, 1493, 1445, 1088, 795, 742, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H3iCl2N2O+H]+: 433.1808, found: 433.1807.
Synthesis of 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1,4-oxazepane, compound 7.
From 4-(2-fluorophenyl)piperidine (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}- 1 ,4-oxazepane (112 mg, 92% yield) was obtained after chromatography EtOAc/MeOH (98:2). IR (ATR): 2936, 2810, 1583, 1490, 1453, 1223, 1138, 1091 , 899, 796, 754, 737, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32FN2O+H]+: 383.2493, found: 383.2492.
Synthesis of 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1,4-oxazepane, compound 8.
From 4-(p-tolyl)piperidine (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane (91 mg, 75% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 3024, 2933, 2808, 1514, 1494, 1452, 1353, 1137, 1090, 1067, 1029, 995, 812, 736, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H35N2O+H]+: 379.2744, found: 379.2739.
Synthesis of 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1,4-oxazepane, compound 9. From 4-(m-tolyl)piperidine (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4- oxazepane (81 mg, 67% yield) was obtained after chromatography EtOAc/MeOH (98:2). IR (ATR): 3025, 2934, 2809, 1607, 1588, 1493, 1452, 1353, 1137, 1089, 1067, 1028, 982, 784, 736, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H35N2O+H]+: 379.2744, found: 379.2741.
Synthesis of 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1,4-oxazepane, compound 10.
From 4-(o-tolyl)piperidine (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane (60 mg, 50% yield) was obtained after chromatography EtOAc/MeOH (98:2). IR (ATR): 2935, 2809, 1603, 1492, 1453, 1260, 1138, 1090, 1067, 1028, 803, 747, 725, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H35N2O+H]+: 379.2744, found: 379.2742.
Synthesis of 3-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound 11.
From 3-(piperidin-4-yl)phenol (96 mg, 0.54 mmol) in MeCN (4 mL), KHCC>3(211 mg, 2.10 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (168 mg, 0.70 mmol) in MeCN (4 mL), 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljphenol (88 mg, 43% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 3027, 2934, 2813, 1599, 1585, 1452, 1353, 1271 , 1155, 1067, 1028, 952, 867, 784, 733, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H33N2O2+H]+: 381.2537, found: 381.2537.
Synthesis of 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 12.
From 4-(4-methoxyphenyl)piperidine (61 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1- yl]methyl}-1 , 4-oxazepane (117 mg, 93% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 2932, 2809, 1611 , 1583, 1512, 1452, 1244, 1177, 1090, 1037, 994, 827, 737, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H35N2O2+H]+: 395.2693, found: 395.2694.
Synthesis of 4-{ 1-[ (4-benzyl- 1, 4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound 13.
From 4-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljphenol (45 mg, 37% yield) was obtained (white powder) after chromatography DCM/MeOH (95:5). IR (ATR): 2940, 2798, 1612, 1515, 1442, 1366, 1270, 1241 , 1180, 1136, 1057, 973, 906, 827, 777, 731 , 697, 592 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32N2O2+H]+: 381.2537, found: 381.2534.
Synthesis of 3-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline, compound 14. To a solution of 3-(piperidin-4-yl)aniline (57 mg, 0.32 mmol) in MeCN (4 mL), KHCC>3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-benzyl-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}aniline (86 mg, 71 % yield) was obtained after chromatography, DCM/MeOH (94:6). IR (ATR): 3339, 2933, 1710, 1597, 1543, 1508, 1446, 1411 , 1311 , 1259, 1235, 1054, 1017, 953, 790,735, 699, 580 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H34N3O+H]+: 380.2696, found: 380.2695.
Synthesis of 4-{ 1-[ (4-benzyl- 1, 4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline, compound 15.
From 4-(piperidin-4-yl)aniline (61 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljaniline (98 mg, 81% yield) was obtained after chromatography DCM/MeOH (96:4). IR (ATR): 3302, 2975, 2937, 1711 , 1594, 1545, 1507, 1413, 1309, 1258, 1219, 1193, 1155, 1052, 1018, 962, 825, 733, 699, 580 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H34N3O+H]+: 380.2696, found: 380.2696.
Synthesis of 3-{ 1-[ (4-benzyl- 1, 4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile, compound 16.
From 3-(piperidin-4-yl)benzonitrile (60 mg, 0.32 mmol) in MeCN (3 mL), KHCO3 (96 mg, 0.96 mmol), Nal and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (3 mL), 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljbenzonitrile (111 mg, 89% yield) was obtained after chromatography DCM/MeOH (90:10). IR (ATR) 3327, 2956, 2852, 1678, 1645, 1502, 1454, 1312, 1189, 1015, 821 , 740, 634 cm’1. HRMS -ESI+ [M+H]+ calcd for [C25H3IN3O+H]+: 390.2545; found 390.2542.
Synthesis of 2-{[4-( 1H-pyrazol- 1-yl)piperidin-1-yl]methyl}-4-benzyl- 1, 4-oxazepane, compound 17.
From 4-(1/7-pyrazol-1-yl)piperidine (48 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl- 1 , 4-oxazepane (39 mg, 34% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 2933, 2886, 2811 , 1510, 1494, 1452, 1396, 1371 , 1308, 1286, 1260, 1137, 1089, 1045, 1028, 963, 741 , 698, 622 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C2IH30N4O+H]+: 355.2492, found: 355.2488. Synthesis of 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol, compound 18. From 4-(3-hydroxyphenyl)piperidine (60 mg, 0.34 mmol) in MeCN (3 mL), KHCC>3 (103 mg, 1.03 mmol), Nal (cat.), and a solution of 4-benzyl-2-chloromethylmorpholine (100 mg, 0.44 mmol) in MeCN (3 mL), 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4- yljphenol (25 mg, 20% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR): 2933, 2886, 2811 , 1510, 1494, 1452, 1396, 1371 , 1308, 1286, 1260, 1137, 1089, 1045, 1028, 963, 741 , 698, 622 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H3oN202+H]+: 367.2380, found: 367.2383.
Synthesis of 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, compound 19.
From 3-(piperidin-4-yl)aniline (60 mg, 0.34 mmol) in MeCN (3 mL), CS2CO3 (289 mg, 0.89 mmol), Nal (cat.), and a solution of 4-benzyl-2-chloromorpholine (59 mg, 0.26 mmol) in MeCN (3 mL), 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline (32 mg, 33% yield) was obtained after chromatography DCM/MeOH (90:10). IR (ATR) 3345, 2918, 2806, 1604, 1453, 1294, 1104, 1027, 866, 781 , 735, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H3oN3OH]+: 366.2545; found 366.2546.
Synthesis of 4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}- 1, 4- oxazepane, compound 20.
From 4-[3-(methylsulfonyl)phenyl]piperidine (57 mg, 0.24 mmol) in MeCN (2 mL), KHCO3 (71 mg, 0.72 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4- oxazepane (74 mg, 0.31 mmol) in MeCN (2.5 mL), 4-benzyl-2-{[4-(3- (methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane (94 mg, 89% yield) was obtained after chromatography DCM/MeOH (90:10). IR (ATR) 3420, 2927, 2811 , 1451 , 1298, 1141 , 1087, 958, 762, 735, 696 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H34N2O3S+H]+: 443.2368; found 443.2364.
Synthesis of 4-benzyl-2-{[4-(pyridin-4-yl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 21.
From 4-(piperidin-4-yl)pyridine (52 mg, 0,32 mmol) in MeCN (3 mL), KHCO3 (96 mg, 0,96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0,42 mmol) in MeCN (3 mL), 4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1 -yl]methyl}-1 ,4- oxazepane (79 mg, 67% yield) was obtained after chromatography DCM/MeOH (90:10). IR (ATR) 3393, 3025, 2934, 2810, 1598, 1451 , 1353, 1135, 1089, 991 , 817, 736, 698, 651 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H3IN3O+H]+: 366.2545; found 366.2538. Synthesis of 4-benzyl-2-{[4-(3-trifluoromethylphenyl) piperidin- 1-yl] methyl}- 1, 4- oxazepane, compound 22.
From 4-[3-(trifluoromethyl)phenyl]piperidine (53 mg, 0.23 mmol) in MeCN (2 mL), KHCO3 (70 mg, 0.69 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4- oxazepane (72 mg, 0,30 mmol) in MeCN (2.5 mL), of 4-benzyl-2-{[4-(3- (trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane (57 mg, 57% yield) was obtained after chromatography DCM/MeOH (90:10). IR (ATR) 2935, 2810, 1449, 1328, 1120, 1072, 914, 800, 738, 699, 675 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H3IF3N2OH]+: 433.2467; found 433.2461.
Synthesis of 4-benzyl-2-{[4-(3-chlorophenyl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 23.
From 4-(3-chlorophenyl)piperidine (63 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}- 1 , 4-oxazepane (85 mg, 66% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR) 2934, 2809, 1596, 1452, 1137, 1088, 1028, 997, 909, 780, 737, 695 cm’ 1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H3ICIN2O+H]+: 399.2198; found 399.2200.
Synthesis of 2-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound 24.
From 2-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljphenol (78 mg, 64% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2940, 2815, 1593, 1454, 1361 , 1279, 1249, 1235, 1108, 1070, 958, 737, 700 cm’ 1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H32N2O2+H]+: 381.2537; found 381.2526.
Synthesis of methyl 3-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate, compound 25.
From methyl 3-(piperidin-4-yl)benzoate (70 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), methyl 3-{1-[(4-benzyl-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}benzoate (97 mg, 72% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2933, 2849, 1720, 1443, 1283, 1241 , 1196, 1095, 980, 752, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C26H34N2O3+H]+: 423.2642; found 423.2633.
Synthesis of 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin- 1-yl) methyl}- 1, 4-oxazepane, compound 26.
From 2-(piperidin-4-yl)pyrimidine (52 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1-yl)methyl}-1 ,4- oxazepane (67 mg, 58% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3387, 2922, 2852, 2811 , 1570, 1559, 1424, 1143, 1088, 1002, 803, 737, 698, 633 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C22H3oN40+H]+: 367.2492; found 367.2491.
Synthesis of 2-{[4-(1H-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1,4- oxazepane, compound 27.
From 2-(piperidin-4-yl)-1/7-benzo[c(]imidazole (64 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4- oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 2-{[4-(1/7-benzo[c(]imidazol-2- yl)piperidin-1 -yl]methyl}-4-benzyl-1 , 4-oxazepane (89 mg, 69% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2933, 1595, 1453, 1361 , 1235, 1074, 953, 897, 803, 737, 699, 645 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for
[C25H32N4O+H]+:405.2649; found 405.2644.
Synthesis of 4-benzyl-2-{[4-(pyridin-3-yl)piperidin- 1-yl]methyl}- 1, 4-oxazepane, compound 28.
From 3-(piperidin-4-yl)pyridine (52 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1 -yl]methyl}-1 ,4- oxazepane (75 mg, 64% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2934, 2811 , 1593, 1453, 1360, 1279, 1235, 1085, 1069, 958, 897, 736, 699, 620 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H3IN3O+H]+: 366.2540; found 366.2538.
Synthesis of 2{[4-( 1H-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl- 1, 4-oxazepane, compound 29.
From 3-(piperidin-4-yl)-1 /7-indole (64 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4- oxazepane (86 mg, 67% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2936, 1593, 1453, 1360, 1278, 1234, 1106, 1068, 1044, 955, 748, 736, 694, 644 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C26H33N3O+H]+: 404.2696; found 404.2697.
Synthesis of 3-{ 1-[ (4-(4-chlorobenzyl)- 1, 4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound 30.
From 3-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-(4-chlorobenzyl)-2-(chloromethyl)-1 ,4-oxazepane (115 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-(4-chlorobenzyl)-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}phenol (99 mg, 74% yield) was obtained after chromatography DCM/MeOH (95:5). IR (ATR) 3124, 2941 , 1615, 1584, 1488, 1407, 1284, 1213, 1166, 1087, 1013, 941 , 817, 790, 704, 634 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H3ICIN2O2+H]+: 415.2147; found 415.2147.
Synthesis of 3-{1-[(4-(4-methylbenzyl)-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound 31.
From 3-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-(4-methylbenzyl)-1 ,4-oxazepane (107 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-(4-methylbenzyl)-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}phenol (98 mg, 78% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3108, 2942, 2876, 1614, 1584, 1488, 1455, 1437, 1408, 1356, 1283, 1213, 1165, 1091 , 940, 799, 705 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H34N2O2+H]+: 395.2693; found 395.2689.
Synthesis of 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine, compound 32.
From 4-(4-chlorophenyl)piperidine (63 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)morpholine (95 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1- yl]methyl}morpholine (95 mg, 77% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3397, 2928, 2805, 1732, 1492, 1452, 1293, 1105, 1090, 1026, 999, 822, 778, 738, 697, 655 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H29CIN2O+H]+: 385.2041 ; found 385.2039.
Synthesis of4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol, compound 33. From 4-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)morpholine (95 mg, 0.42 mmol) in MeCN (4 mL), 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol (92 mg, 78% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2945, 2818, 2780, 1611 , 1513, 1465, 1442, 1266, 1239, 1110, 1022, 909, 828, 746, 697 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H3oN202+H]+ :367.2380; found 367.2378.
Synthesis of 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine, compound 34.
From 4-phenylpiperidine (52 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)morpholine (95 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine (87 mg, 77% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3028, 2944, 2779, 1610, 1513, 1452, 1267, 1239, 1104, 1023, 996, 827, 805, 746, 697 cm-1. HRMS- ESI+ m/z [M+H]+ calcd for [C23H3oN20+H]+: 351.2431 ; found 351.2427.
Synthesis of 4-benzyl-2-{[4-(3-(trifluoromethyl)phenyl)piperidin-1-yl]methyl}morpholine, compound 35.
From 4-(3-(trifluoromethyl)phenyl)piperidine (73 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2- (chloromethyl)morpholine (95 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(3- trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine (100 mg, 75% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2933, 2854, 2810, 1738, 1611 , 1513, 1450, 1328, 1245, 1161 , 1111 , 1026, 911 , 800, 746, 675, 654 cm’1. HRMS- ESI+ m/z [M+H]+ calcd for [C24H329N2F3+H]+: 419.2305; found 419.2300.
Synthesis of 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine, compound 36.
From 4-(o-tolyl)piperidine (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)morpholine (95 mg, 0.42 mmol) in MeCN (4 mL), 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine (84 mg, 72% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3380, 2920, 2855, 2806, 2457, 1452, 1291 , 1110, 1074, 1035, 955, 826, 749, 699, 674 cm’1. HRMS- ESI+ m/z [M+H]+ calcd for [C24H32N2O+H]+: 365.2587; found 365.2585.
Synthesis of 2-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline, compound 37. From 2-(piperidin-4-yl)aniline (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljaniline (65 mg, 54% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3061 , 3024, 2935, 2810, 1621 , 1579, 1494, 1453, 1290, 1255, 1136, 1106, 1089, 994, 981 , 926, 810, 742, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H33N3O+H]+: 380.2696; found 380.2701.
Synthesis of 5-{ 1-[ (4-benzyl- 1, 4-oxazepan-2-yl) methyl]piperidin-4-yl}-2-methylphenol, compound 38.
From 2-methyl-5-(piperidin-4-yl)phenol (61 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 5-{1-[(4-benzyl-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}-2-methylphenol (86 mg, 68% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2922, 2852, 1732,1617,
1590,1452,1420,1374,1290, 1242, 1119, 1107, 1074, 952, 867, 735, 698, 640 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H34N2O2+H]+:395.2693; found 395.2697.
Synthesis of 5-{ 1-[ (4-benzyl- 1, 4-oxazepan-2-yl) methyl]piperidin-4-yl}-2-fluorophenol, compound 39.
From 2-fluoro-5-(piperidin-4-yl)phenol (62 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 ,4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 5-{1-[(4-benzyl-1 ,4-oxazepan-2- yl)methyl]piperidin-4-yl}-2-fluorophenol (82 mg, 65% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3670, 2936, 1602, 1528, 1514, 1494, 1451 , 1431 , 1375, 1353, 1297, 1275, 1241 , 1106, 906, 868, 807, 769, 734, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H3iN2O2F+H]+:399.2442; found 399.2444.
Synthesis of 3-{ 1-[ (4-(4-methoxybenzyl)-1 , 4-oxazepan-2-yl)methyl]piperidin-4- yl}phenol, compound 40.
From 3-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-(4-methoxybenzyl)-1 ,4- oxazepane (113 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-(4-methoxybenzyl)-1 ,4- oxazepan-2-yl)methyl]piperidin-4-yl}phenol (95 mg, 72% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3670, 2923, 1610, 1584, 1511 , 1453, 1243, 1170, 1151 , 1107, 1077, 1033, 952, 855, 834, 811 , 783, 699 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C25H34N2O3+H]+: 411.2642; found 411.2646.
Synthesis of 3-{ 1-[ (4-(3, 4-dichlorobenzyl)-1 , 4-oxazepan-2-yl)methyl]piperidin-4- yl}phenol, compound 41.
From 3-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-(3,4-dichlorobenzyl)-1 ,4- oxazepane (130 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-(3,4-dichlorobenzyl)-1 ,4- oxazepan-2-yl)methyl]piperidin-4-yl}phenol (111 mg, 77% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3444, 2937, 1612, 1583, 1512, 1483, 1454, 1391 , 1350, 1291 , 1247, 1169, 1131 , 1108, 1078, 1033, 997, 976, 954, 867, 830, 818, 783, 705, 669 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C24H3oN202Cl2+H]+: 449.1757; found 449.1759.
Synthesis of 3-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound
42.
From 3-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 ,4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljphenol (76 mg, 78% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2936, 2806, 1613, 1598, 1583, 1453, 1390, 1349, 1277, 1248, 1131 , 1107, 1077, 953, 868, 783, 700, 669 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [Ci8H28N2O2+H]+: 305.2224; found 305.2225.
Synthesis of 3-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline, compound
43.
From 3-(piperidin-4-yl)aniline (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 ,4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljaniline (71 mg, 74% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2935, 2799, 1605, 1585, 1459, 1391 , 1348, 1293, 1248, 1131 , 1106, 1078, 953, 866, 783, 699 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [Ci8H29N3O+H]+: 304.2383; found 304.2386.
Synthesis of 4-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, compound 44. From 4-(piperidin-4-yl)phenol (57 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 ,4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljphenol (80 mg, 82% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2939, 2917, 2880, 2846, 2826, 2797, 1613, 1587, 1517, 1453, 1387, 1247, 1193, 1168, 1146, 1128, 1096, 848, 831 , 818, 794, 779, 760 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [Ci8H28N2O2+H]+: 305.2224; found 305.2230.
Synthesis of 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl- 1, 4-oxazepane, compound 45.
From 4-(4-methoxyphenyl)piperidine (61 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 , 4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4- methyl-1 , 4-oxazepane (86 mg, 85% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3395, 2935, 1611 , 1512, 1244, 1178, 1131 , 1107, 1094, 1034, 830, 716 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [Ci9H3oN202+H]+: 319.2380; found 319.2384.
Synthesis of 4-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline, compound 46.
From 4-(piperidin-4-yl)aniline (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 , 4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4- yljaniline (71 mg, 74% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3416, 3313, 3187, 2936, 2797, 1639, 1614, 1517, 1454, 1281 , 1251 , 1131 , 1106, 1096, 1069, 982, 857, 822, 775 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [CI8H29N3O+H]+: 304.2383; found 304.2388.
Synthesis of 4-methyl-2-{[4-(o-tolyl) pi peridi n-1-yl]methyl}-1, 4-oxazepane, compound 47. From 4-(o-tolyl)piperidine (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 , 4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 4-methyl-2-{[4-(o-tolyl)piperidin-1 -yl]methyl}-1 , 4-oxazepane (71 mg, 79% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2935, 2792, 1460, 1133, 1107, 1094, 749, 724 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [CI9H30N2O+H]+: 303.2431 ; found 303.2434. Synthesis of 2-{[4-(3-Methoxyphenyl)piperidin-1-yl]methyl}-4-methyl- 1, 4-oxazepane, compound 48.
From 4-(3-methoxyphenyl)piperidine (61 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 2-(chloromethyl)-4-methyl-1 , 4-oxazepane (69 mg, 0.42 mmol) in MeCN (4 mL), 2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4- methyl-1 , 4-oxazepane (84 mg, 83% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 3392, 2936, 1731 , 1608, 1601 , 1582, 1484, 1464, 1452, 1262, 1241 , 1152, 1131 , 1107, 1094, 1043, 868, 781 , 752 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [Ci9H3oN202+H]+: 319.2380; found 319.2383.
Synthesis of 3-{ 1-[ (4-benzyl- 1, 4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol, compound 49.
From 3-(pyrrolidin-3-yl)phenol (52 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)-1 , 4-oxazepane (100 mg, 0.42 mmol) in MeCN (4 mL), 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]pyrrolidin-3- yljphenol (79 mg, 68% yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2917, 2834, 1599, 1585, 1452, 1375, 1353, 1279, 1241 , 1148, 1072, 961 , 865, 783, 737, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H30N2O2+H]+: 367.2380; found 367.2387.
Synthesis of 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, compound 50.
From 4-(piperidin-4-yl)aniline (56 mg, 0.32 mmol) in MeCN (4 mL), KHCO3 (96 mg, 0.96 mmol), Nal (cat.), and a solution of 4-benzyl-2-(chloromethyl)morpholine (95 mg, 0.42 mmol) in MeCN (4 mL), 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline (82 mg, 71 % yield) was obtained after chromatography DCM/MeOH (94:6). IR (ATR) 2920, 2851 , 2808, 1731 , 1615, 1517, 1452, 1289, 1248, 1107, 972, 821 , 773, 738, 698 cm’1. HRMS-ESI+ m/z [M+H]+ calcd for [C23H3IN3O+H]+: 366.2540; found 366.2543.
G9a inhibitors were serially diluted between 5 nM and 0.001 nM in 100 % DMSO (Sigma, St. Louis, USA). Then, respective concentrations were subsequently diluted in MilliQ purified water to reach a final concentration ranging between 50 and 0.001 pM in 1 % DMSO in well. In vitro determination of G9a (EHMT2) inhibition activity
The following fluorescent assay was used for determination of the G9a (also called EHMT2, the euchromatic histone methyltransferase 2) inhibition activity (IC50). G9a activity was measured using the G9a Chemiluminescent Assay Kit (Catalog # 52001 L, BPS Bioscience, San Diego, CA, USA), following manufacturer’s instructions. The experiment was run per triplicate and are shown in Table 1 below:
Table 1. IC50 of each G9a inhibitor compound. In vitro determination of GLP (EHMT1) inhibition activity
Since, G9a and G9a-like (GLP, also known as EHMT1, euchromatin histone lysine methyltransferase 1) are members of the Suv39h subgroup of SET domain-containing molecules, and together they are the key HKMTs for H3K9me1 and H3K9me2. Then, to confirm the selectivity against G9a we performed the following fluorescent assay was used for determination of the GLP inhibition activity (IC50), with substrate and comparative control compound UNC0638, which corresponds to 2-cyclohexyl-/V-(1- isopropylpiperidin-4-yl)-6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-amine: which is a well-established inhibitor with a high potency of G9a and GLP. Unfortunately, the binding to GLP is undesirable in the context of the invention because it gives rise to cytotoxicity. Protocol:
GLP activity was measured using the GLP Chemiluminescent Assay Kit (Catalog # The experiment was run per triplicate and are provided in Table 2 below.
Table 2.
Table 2 shows percentage of inhibition of GLP from different assayed compounds (three different experiments in triplicate). Of note, compound of the invention Compound 11 does not significantly inhibit the GLP target in comparison UNC0638. This means a great improvement with respect to the reference treatment, as no toxicity side-effect would be provided by the molecules of the invention.
Therefore, it is shown that the compounds of the inventions are not only potent inhibitors of G9a, but also highly selective of this target and safe.
Western blot: H3K9me2 main epigenetic mark modulated by G9a (target engagement)
Histones extraction, and Western Blotting
To confirm the target engagement after G9a inhibition, we determined the ratio of H3 K9me2/H3 total N2 (WT) and CL2006 animals obtained from the Genetics Center, CGC (University of Minnesota, MN, USA) were incubated in distilled water with vehicle (DMSO 1 %), or 0.1 pM of Compound 11 our G9a inhibitor in distilled water with (DMSO 1 %) in a 96-well plate format as already mentioned. For chronic treatment, four-day-old animals were collected with M9 buffer. Histone extraction was performed following the manufacturer’s instructions (EpiQuik Total Histone Extraction HT Kit, EpiGentek, #OP- 0007-192). The samples were resolved in a 14% SDS-gel, as previously described. To capture chemiluminescence signals were used Amersham Imager 680 and Western blot quantifications were performed using ImageLab software (Bio-Rad). Immunoblots were probed with anti-H3 K9me2 (1 : 1000) (Epigentek, #A-4035), and anti-H3 total signaling, #9715).
Table 3. Quantification of the total H3K9me/H3 ratio after the pharmacological treatments. Values represented are mean ± Standard error of the mean (SEM); n = 3 with at least 200 worms in each group. Statistical analysis: Unpaired t-test: N2 (WT) vs. CL2006 Vehicle, *p<0.05; One-Way ANOVA: Dunnett’s post-hoc test: CL2006 Vehicle vs. Compound 11 , and UNC0638, *p<0.05.
Table 3 shows that these compounds directly target G9a. First, we compared the ratio H3K9me2/H3 total levels in the N2 (WT) strain compared to the AD transgenic strain (CL2006). The ratios were obtained dividing the H3K9me2 levels between total amount of histone 3 multiplied by 100 for each group separately. Interestingly, the CL2006 presented higher levels of H3K9me2, confirming the link between this repressive mark and AD pathology. As for Compound 11 , we observed that the ratio H3K9me2/H3 total levels after each pharmacological intervention were statistically reduced at the dose of 0.1 pM, unlike the reference compound UNC0638.
In vitro determination of hERG activity (safety test)
Determination of hERG activity was performed because blocking hERG channels represents a major therapeutic challenge in drug discovery, as it is an important safety issue.
CHO cells stably expressing hERG channels (Millipore) were cultured in F12 HAM medium supplemented with 10% FBS and 400 pg/L Geneticin. The extracellular Ringer’s solution consisted of (in mM): 2 CaCh, 1 MgCh, 10 HEPES, 4 KCI, 145 NaCI, 10 glucose, pH 7.4, 305 mOsm. The intracellular Ringer’s solution consisted of (in mM): 5.37 CaCh, 1.75 MgCh, 31.25/10 KOH/EGTA, 10 HEPES, 210 KCI, pH 7.2, 295 mOsm. 4 mM Na2- ATP was added to intracellular Ringer’s solution shortly before use. Whole-cell currents were measured with a QPatch system (Sophion) in response to continuously executed voltage protocols as per manufacturer’s recommendations. Upon onset of the voltage protocol, cells were maintained at a holding potential (Vh) of -80 mV, then clamped briefly to -50 mV (20 ms), subsequently depolarized to 20 mV for 4800 ms, and finally repolarized to -50 mV for 5000 ms, at which potential the peak outward tail current was measured. Finally, the voltage returned to Vh for 3100 ms. Thus, voltage protocols were repeated each 15 seconds. For each cell, extracellular solution was applied before increasing concentrations of the tested compound.
Table 4. The table summarizes the IC50 hERG obtained for assayed compounds.
Table 4 shows hERG inhibitory activity compared to the control (haloperidol).
Since blocking hERG channels represent a major therapeutic challenge in drug discovery, we evaluated hERG inhibitory activity. As it is shown in Table 4, the compounds of the invention provided much higher IC50 values than haloperidol. The higher the value, the lower the affinity, and, therefore, the compounds are safer.
Therefore, it is confirmed that the high potency, even at high concentrations, does not negatively affect to the safety of the molecules of the invention.
In vitro determination of human plasma protein binding (bioavai labi I ity test)
In drug discovery it is of interest that the binding to plasma proteins is low because there will be more drug available in plasma that can exert its effect. However, if it is too low it will be eliminated very quickly, so what is sought is a balance.
Plasma from Seralab was employed in the assay. Briefly, the assay was carried out by employing Rapid Equilibrium Dialysis (RED) from Thermo Scientific. Compound 11 was dissolved at 5 pM in plasma and added to the corresponding insert of the RED device.
Dialysis buffer was added to the corresponding insert of the RED device. Plate was incubated for 4h at 37°C. After the incubation period 50 pL aliquots of each chamber were transferred to empty vials. 50 pL of dialysis buffer were added to the plasma samples and 50 pL of plasma were added to the buffer samples. 300 pL of acetonitrile were added to all the samples and centrifuged at 4000 rpm. 100 pL aliquots of the supernatants were transferred to a LC analysis plate. Samples were analyzed in a UPLC/MS/MS. Stationary phase: ACQUITY BEH C18 1.7pm 2.1x50mm (Waters)/ ACQUITY HSST3 1.8pm 2.1x100mm (Waters). Gradient: A (H2O + 0.1 % Formic acid); B (ACN + 0.1% Formic acid); Flow: 0.6mL/min.
TABLE A t(min) A (%) B (%)
0 100 0
0,1 100 0
1 0 100
2 0 100
2,1 100 0
2.5 100 0
The chromatographic equipment employed was an ACQUITY LIPLC I Xevo TQD System. Compound concentrations were calculated from the MS peak areas.
Table 5. Percentage of Compound 11 of human plasma protein biding.
Table 5 shows that Compound 11 has a good plasma protein binding with free compound value lower than 99%, but higher than the reference compound, Phenytoin. Phenytoin is chosen as a positive control as it is extensively unbound fraction in plasma about 20% in different animal species.
In vitro determination of plasma stability in human plasma
Human plasma pooled from healthy donors extracted in citrate tubes was employed in the assay. Briefly, plates containing 5pM compounds in plasma (total volume: 100pL) were incubated at 37°C at the different times (0, 60, 120 and 360 min). Then, 300pl Acetonitrile were added for precipitating plasma protein, and the plate was centrifuged at 4000 g for 60 min at 4°C. Supernatant was taken and analyzed by UPLC/MS/MS for sample quantification. Stationary phase: ACQUITY BEH C18 1.7pm 2.1x50mm (Waters)//ACQUITY HSST3 1.8pm 2.1x100mm (Waters). Mobile phase: 0.1 % Formic acid water/0.1 % formic acid in acetonitrile.
TABLE B: Gradient
Time Water Acetonitrile 0 100% 0% 0.1 100% 0% 1 0% 100% 2 0% 100% 2.1 100% 0% 2.5 100% 0%
Flow: 0.6 ml/min
The chromatographic equipment employed was an LIPLC QSM Waters Acquity.
Compound concentrations were calculated from the MS peak areas.
Table 6. Remaining percentage of Compound 11 in plasma at the different times tested.
This study evaluates the ability of Compound 11 to degrade in plasma. Table 6 shows that after 6 hours, Compound 11 still remains in at a remarkable high amount, which is indicative of its plasma stability.
In vitro determination of microsomal stability at human microsomes
Microsomal stability, which is widely used to determine the likely degree of primary metabolic clearance in the liver, was assessed in human recombinant microsomes.
The human microsomes from different suppliers, 1A2 was purchased from Corning reference 456203, 2C9 was purchased from Tebu Bio reference CYP/EZ037, and 2D6 was purchased from Corning reference 456217. The compound was incubated at 37 °C with the microsomes in a 50 mM phosphate buffer (pH = 7.4) containing 30 mM MgCh, 10 mM NADP, 100 mM glucose-6-phosphate, and 40 U/rnL glucose-6-phosphate- dehydrogenase. Samples (75 pL) were taken from each well at 0, 10, 20, 40, and 60 min and transferred to a plate containing 4 °C 75 pL of acetonitrile and 30 pL of 0.5% formic acid in water were added for improving the chromatographic conditions. The plate was centrifuged (46,000g, 30 min) and supernatants were taken and analyzed by an ultraperformance liquid chromatograph-tandem mass spectrometer (Xevo-TQD, Waters) by employing a BEH C18 column and an isocratic gradient of 0.1% formic acid in water: 0.1 % formic acid acetonitrile (60:40).
The metabolic stability of the compounds was calculated from the logarithm of the remaining compounds at each of the time points studied.
Table 7. The table summarizes the stability parameters obtained for compound Compound 11.
Table 7 shows a percentage remanent at 60 minutes. Of note, Compound 11 demonstrated an 40.6 % remanent at 60 minutes in comparison of Testosterone, which is a control for this technique. Likewise, the t1/2 (min) column indicates how long (in minutes) it took for the compound to degrade by 50%. In this case, the half-life is 45.62 minutes, which coincides with the remaining percentage, since 60% had already degraded after 60 minutes. The Compound 11 had intermediate levels. The lower the half-life, the more the compound is metabolized. This value is calculated with an adjustment of the remaining % In at each of the times studied versus time. The antilogarithm of the slope of that fit is the half-life value. Moreover, the third value (Clint) indicates the intrinsic clearance. The higher this value is, it indicates that the compound degrades more quickly, so it would be interesting for the intrinsic clearance values to be as low as possible. Again, the value we obtained tells us that Compound 11 had an intermediate metabolism rate. This value is calculated taking into account the number of microsomes used in the assay, the obtained half-life value and the incubated volume, so it is a value that depends on the half-life calculation. In fact, in vitro CLint is calculated from HLM, recombinant enzymes and hepatocytes using Eqs. (4)-(6), respectively.
All these models assume that there is no permeability limitation within the liver and drug entry into liver is perfusion rate limited.
In conclusion, the C t value is lower in Compound 11 in comparison with Testosterone, indicating an excellent stability. Testosterone is chosen as a positive control as it is extensively metabolized by several enzymes and in a characteristic way by different species, making it a good positive control for metabolic activity.
Parallel Artificial Membrane Permeation Assays - Blood-Brain Barrier (PAMPA- BBB)
To evaluate the brain penetration of the different compounds, a parallel artificial membrane permeation assay for blood-brain barrier was used, following the method described by Di et al. (2003).
The in vitro permeability (Pe) of fourteen commercial drugs through lipid extract of porcine brain membrane together with the test compounds were determined.
Commercial drugs and assayed compounds were tested using a mixture of PBS:ETOH (70:30).
Assay validation was made by comparing the experimental permeability with the reported values of the commercial drugs by bibliography and lineal correlation between experimental and reported permeability of the fourteen commercial drugs using the parallel artificial membrane permeation assay was evaluated (y=1.623x-1.332; R2=0,9426). Table 8 shows permeability results from the different commercial and assayed compounds (three different experiments in triplicate) and predictive penetration in the CNS.
Table 8. Permeability (Pe 10'6 cm s-1) in the PAMPA-BBB assay of tested compounds and predictive penetration in the CNS. Di, L.; Kerns, E. H.; Fan, K.; McConnell, O. J.; CarTer, G. T. High throughput artificial membrane permeability assay for blood-brain barrier. Eur. J. Med. Chem. 2003, 38, 223- 232.
*Spectral absorbance 210-250 nm All the compounds of the invention showed a remarkable improved ability to penetrate BBB, when compared with the reference compound UNC0638.
In vitro evaluation in a preliminary security panel
The objective of this study was to screen the potential inhibition of the compounds using recombinant human cytochrome P450 (CYP450) enzymes. CYP450 is a major drugmetabolizing enzyme system mainly distributed in liver and involved in the metabolism of many endogenous substance, and exogenous compounds such as drugs, toxicants, carcinogens, and procarcinogens). There are different families of CYP450 such as (CYP1A2, CYP2C9 and CYP2D6), which are mainly expressed in liver, small intestine, placenta and kidney, and they are involved in the metabolism of drugs accounting for 80% were tested using probe substrates with fluorescent detection.
Incubations were conducted in a 200 pL volume in 96 well microtiter plates (COSTAR 3915). Addition of cofactor-buffer mixture (KH2PO4 buffer, 1.3 mM NADP, 3.3 mM MgCh, 3.3 mM Glucose-6-phosphate and 0.4 U/rnL Glucose-6-phosphate Dehydrogenase), supersomes control, standard inhibitors (Furafyline, Tranylcypromine, Sulfaphenazole and Quinidine; from Sigma Aldrich) previously diluted and compounds to plates were carried out by a liquid handling station (Zephyr Caliper). The plate was then preincubated at 37°C for 5 min, and the reaction initiated by the addition of pre-warmed enzyme/substrate (E/S) mix. The E/S mix contained buffer (KH2PO4), c-DNA-expressed P450 in insect cell microsomes, substrate (3-cyano-7-ethoxycoumarin (CEC for Sigma Aldrich reference LIC455) for CYP1A2, 7-methoxy-4-(trifluoromethyl)coumarin (7-MFC from Sigma Aldrich reference T3165) for CYP2C9, and 3-[2-(/V, /V-Diethyl-A/- methylammonium)ethyl]-7-methoxy-4-methylcoumarin (AMMC from Corning reference 451705) for CYP2D6) and other components to give the final assay concentrations in a reaction volume of 200 pL. Reactions were terminated after various times (a specific time for each cytochrome) by addition of STOP solution (ACN/TrisHCI 0.5M 80:20). Fluorescence per well was measured using a fluorescence plate reader (Tecan M1000 infinite pro) and percentage of inhibition was calculated.
Table 9. Results of inhibition of Compound 11 Table 9 shows that Compound 11 has low inhibitory activity at 10 pM at the different cytochromes evaluated. Taking into account the nanomolar range of G9a inhibition values of Compound 11 it is not expected that at therapeutic doses the compound is going to interfere with the CYP450 mediated metabolism of other drugs. In conclusion, we demonstrated that our G9a inhibitors are safe, avoiding metabolism interactions with other drugs.
In vivo results in C. elegans
Statistics analysis
The statistical analysis was conducted using GraphPad Prism version 9.2 statistical software. Group size may differ depending on power analysis and expertise of the authors. Shapiro-Wilk test to verify data normality for all groups. Data were expressed as the mean ± Standard Error of the Mean (SEM). For normally distributed data, means were compared in One-Way or Two-Way ANOVA analysis of variance, (ANOVA), followed by Tukey’s post-hoc analysis. Comparison between groups was also performed by a two-tailed Student’s t-test for independent samples. In contrast, the Mann-Whitney or Kruskal-Wallis test followed by Dunn’s post-hoc analysis was used for non-normally distributed data. Statistical significance was considered when p-values were <0.05. T. For behavioral tests, a blinded analysis was performed.
C. elegans model
The WT C. elegans strain (N2), and the transgenic CL2006 (dvls2 [pCL12(unc-54/human Ap peptide 1-42 minigene)+rol-6(su1006)]) provided by the C. elegans Genetic Center were used. Standard methods were used for culturing and observing C. elegans. N2 were propagated at 20°C, while CL2006 worms were maintained at 16°C in a temperature-controlled incubator on solid nematode growth medium (NGM) seeded with Escherichia coli (E. coli) OP50 (Carolina Biological) strain as a food source.
Pharmacological treatment, and locomotion assay
Locomotion assay of G9a inhibitors was assessed to obtain dose-response profile, evaluating the impact of the pharmacology treatment in motor dysfunction presented by the transgenic strains CL2006. Worms were grown with continuous shaking at 180 rpm at 20 °C for 4 days. Each well contained a final volume of 60 pL, comprising 25- 30 animals in the larva 1 (L1) stage diluted in S-medium solution, G9a inhibitor at the appropriate dose (means that the working solution is 2.4 fold more concentrated than the final concentration in the well), and OP50 (Carolina Biological) inactivated by freezethaw cycles suspended in S-medium complete solution to a final OD595 of 0.9 measured in the microplate reader. On day 5 of age, worms were transferred from the 96-well plates onto an unseeded NGM plate for 45 minutes before starting the trial, allowing the plates to dry.
Then, locomotion assays were performed in 30 mm NGM plates, in which the whole surface of the plate was covered by OP50 (Carolina Biological, item: 155073). 5 to 10 adult nematodes were placed in the center of a circle (with 1 cm of diameter) in the seeded 30 mm NGM plates. After 1 min, the number of animals remaining inside the circle was scored as a locomotor defect (“LD”). Motor behavior assays were run in triplicates (n=3), with a total of at least 100 animals tested per compound concentration. Motor index establishes a score where treated animals showing the same motor defect as CL2006 have an index of 0%. By contrast, when animals improve motor behavior comparable to the WT (N2) they get an index of 100%. All the results shown in the figures are calculated using the following formula:
. . . . , ,n/ , (LD CL2006 vehicle - LD CL2006 drug) . > _
Motor index (%) = 1 - — x 100
' ' (LD CL2006 vehicle - LD WT vehicle)
LD = locomotion defective
Table 10. Values represented are mean ± Standard error of the mean (SEM); n = 3 with at least 90-100 worms CL2006 in each group. Statistical analysis: One-Way ANOVA, followed by Dunnett post-hoc analysis. 0 pM vs. UNC0638 treatment:***p<0.001 ; ****p<0.0001. 0 pM vs. Compound 11 : ####p<0.0001. One-Way ANOVA, followed by Fisher’s LSD post-hoc analysis. 0.01 pM UNC0638 Vs. 0.01 pM C: $$p<0.01 ; 0.001 pM UNC0638 Vs. 0.001 pM C: $p<0.05.
Table 10 shows that the Compound 11 motor index decreased progressively with the doses tested. Strikingly, at low doses (0.01 pM, and 0.001 pM) Compound 11 showed a statistically significantly greater restoration of motor index at an order of magnitude lower concentration (approximately 30%) in CL2006 compared to UNC0638. Thus, Compound 11 ameliorates the defective locomotion caused by paralysis in the CL2006 worm due to age and Ap plaques, suggesting the G9a inhibitions as a therapeutic strategy for AD.
Thioflavin-S staining AP aggregation
After 5 days of treatment, adult CL2006 C. elegans were fixed in 4% Paraformaldehyde/Phosphate-buffered saline (PBS) (pH 7.5), for 24 hours at 4°C. Then, worms were permeabilized in 5% fresh p-mercaptoethanol, 1% Triton X-100, 125 mm Tris (pH 7.5), at 37°C for another 24 hours. On the last day, nematodes were stained with 0.125% Thioflavin-S (ThS) (Sigma, CAS# 1326-12-1) in 50% ethanol (EtOH) for 2 min, destained in 50% EtOH for 2 min, washed 3 times with PBS.
To prepare the glass slide for microscopy, approximately 10 pL volume was transferred on a droplet of Fluoromount G (Electron Microscopy Sciences, CAT#17984-25). Fluorescence images were acquired using a 20 A~ objective of a fluorescence microscope. Ap in the head region of worms were quantified by counting the number of ThS positive spots using Imaged and were expressed as Ap deposits/anterior area. Ap aggregates were scored by an investigator blinded to G9a inhibitors treatments.
Table 11. Quantification of Thioflavin S-positive particles in the head region of CL2006 strain. Values represented are mean ± Standard error of the mean (SEM); n = 3 with 25 worms in each group. Statistical analysis: One-Way ANOVA, followed by Dunnett post- hoc analysis. Vehicle vs. Compound 11 : ***p<0.001 ; ****p<0.0001. Statistical analysis: Unpaired t-test. UNC0638 0.1 pM vs. Compound 11 0.1 pM: ## p<0.001. Table 11 shows that the Compound 11 reduced Ap aggregation by approximately 45% compared to the vehicle group, being more effective in reducing Ap aggregation than UNC0638, which only reduced Ap deposition by approximately 26%.
Activity on mouse model of Alzheimer disease
Statistics analysis
The statistical analysis was conducted using GraphPad Prism version 9.2 statistical software. Group size may differ depending on power analysis and expertise of the authors. Shapiro-Wilk test to verify data normality for all groups. Data were expressed as the mean ± Standard Error of the Mean (SEM). For normally distributed data, means were compared in One-Way or Two-Way ANOVA analysis of variance, (ANOVA), followed by Tukey’s post-hoc analysis. Comparison between groups was also performed by a two-tailed Student’s t-test for independent samples. In contrast, the Mann-Whitney or Kruskal-Wallis test followed by Dunn’s post-hoc analysis was used for non-normally distributed data. Statistical significance was considered when p-values were <0.05. T. For behavioral tests, a blinded analysis was performed.
Animal model
Senescence-accelerated mice prone 8 (SAMP8) is a model of age-related neurodegeneration underlying LOAD. This model, established through phenotypic selection from AKR/J mice, presents AD-like cognitive symptoms and behavioural abnormalities, including anxiety-like behaviour and depression. Additionally, SAMP8 brains are characterized by pathological signatures of AD, including neuroinflammation, synaptic deficits, oxidative stress and aberrant epigenetic dysregulation (Grinan-Ferre C, Corpas R, Puigoriol-lllamola D, Palomera-Avalos V, Sanfeliu C, Pallas M. Understanding Epigenetics in the Neurodegeneration of Alzheimer's Disease: SAMP8 Mouse Model. J Alzheimers Dis. 2018;62(3):943-963. doi: 10.3233/JAD-170664. PMID: 29562529; PMCID: PMC5870033.).
Treatment
Animals were treated for 15 days with vehicle (control, (20% w/v, (2-hydroxypropyl)-p- cyclodextrin)) or the Compound 11 compound via I.P.. The test compound was dissolved in 20% w/v, (2-hydroxypropyl)-p-cyclodextrin and volume of injection was calculated according to the animal weight to reach the precise daily dose. A freshly made weekly replaces the drinking solution. Animals had free access to food and water and were kept under standard temperature conditions (22 ± 2°C) and 12h:12h light-dark cycles (300 lux/0 lux). After the treatment period, cognitive tests were performed on the animals. After 15 days of dayli treatment, mice were studied in the behavioural tests.
Behavioral tests
The in vivo models for assessing the efficacy of a test compound in learning and memory impairment were based on Novel Object Recognition Test; NORT, and Object Location Test; OLT.
Novel Object Recognition Test (NORT)
Mice were place 90-degree, two-arm, 25-cm-long, 20-cm-high maze of black polyvinyl chloride. Light intensity in the middle of the field was 30 lux. First, mice were individually habituated to the apparatus for 10 min per day for 3 days. On day 4, the animals were allowed to freely explore two identical objects (A and A or B and B) placed at the end of each arm for a 10 min acquisition trial (first trial-familiarization). Then, a 10-min retention trial (second trial) was carried out 2 h (short-term memory) or 24 h (long-term memory) later. During the Short-term memory retention test, the times that the animal spent exploring the new object (TN) and the old object (TO) was recorded. Twenty-four hours after the acquisition trial, the mice were tested again, with a new object and an object identical to the new one in the previous trial (A and C, or B and C). TN and TO were measured from the video recordings from each trial session. A Discrimination index (DI) was defined as (TN-TO)/(TN+TO). The maze, the surface, and the objects were cleaned with 70% ethanol between the animals’ trials to eliminate olfactory cues.
The learning and memory paradigm is based on the spontaneous exploratory activity of rodents and does not involve rule learning or reinforcement. The object recognition paradigm has been shown to be sensitive to the effects of aging and cholinergic dysfunction, among others (Neurosci. Lett. 1994, vol. 170, pp 117-120; Pharmacol. Biochem. Behav. 1996, vol. 35 53, pp. 277- 283). This model has been adapted to mice and validated using pharmacological agents (Front. Biosci. (Schol. Ed.) 2015, vol. 7, pp 10-29). Table 12. NORT evaluation by DI after 2h and 24h. Values presented are the mean ± SEM; (SAMP8 Control n = 10, and SAMP8 Compound 11 (5mg/Kg) n = 10); Statistics analysis: Student’s t-test analysis; *p<0.05.
Table 12 above shows working memory results by using Novel object recognition test after G9a inhibition treatment with our candidate Compound 11. Interestingly, we showed a significant improvement in SAMP8 treated with Compound 11 in comparison with SAMP8 Control in short- and long-term memories, suggesting the G9a participation of cognitive impairment in Alzheimer’s Disease.
Object Location Test (OLT)
OLT was performed using a white plywood apparatus (50 x 50 x 25 cm), in which three walls were white and one was black. On the first day, animals were just habituated to the empty open field arena for 10 minutes. On the second day, two objects were placed in front of the black wall, equidistant from each other and the wall. The objects were 10 cm high and identical. The animals were placed into the open field arena and allowed to explore both objects and surroundings for 10 minutes. Afterward, animals were returned to their home cages, and the OLT apparatus was cleaned with 70% ethanol. On the third day, one object was moved in front of the opposite white wall to test spatial memory. Trials were recorded using a camera mounted above the open field area, and the total exploration time was determined by scoring TN and TO. DI was calculated, defined as (TN-TO)/(TN+TO). The OLT evaluates the spontaneous tendency of rodents to spend more time exploring a novel object location than a familiar object location and recognizing when an object has been relocated.
Table 13. OLT evaluation by DI after changing the location of one object. Values presented are the mean ± SEM; (SAMP8 Control n = 10, and SAMP8 Compound 11 (5mg/Kg) n = 10); Statistics analysis: Student’s t-test analysis; *p<0.05.
|~Group | Number of mice | DI (-1 ,1) 1
Table 13 shows spatial memory results by using Object location after G9a inhibition treatment with our candidate Compound 11. Interestingly, we showed a significant improvement in SAMP8 treated with Compound 11 in comparison with SAMP8 Control in the discrimination index (DI).
Brain tissue dissection
After behavioral tests, SAMP8 mice were euthanized by cervical dislocation. Brains were immediately removed from the skull. Cortex and hippocampus were then isolated and frozen on powdered dry ice. They were maintained at -80 °C for biochemical experiments. For the Golgi staining protocol, see the procedure in the section “Spine density and Golgi staining protocol”.
Histones extraction, and Western Blotting
Histone extraction was performed following the manufacturer’s instructions using hippocampal tissue from each experimental group (EpiQuik Total Histone Extraction HT Kit, EpiGentek, #GP-0007-192). The samples were resolved in a 14% SDS-gel, as previously described. [12] To capture chemiluminescence signals were used Amersham Imager 680 and Western blot quantifications were performed using ImageLab software (Bio-Rad). Immunoblots were probed with anti-H3 K9me2 (1 :1000) (Epigentek, #A- 4035), and anti-H3 total signaling, #9715).
Table 14. Quantification of H3K9me2 in SAMP8 and SAMP8 mice treated with Compound 11. (Values presented are mean ± SEM; (SAMP8 Control n = 6, and SAMP8 Compound 11 (5mg/Kg) n = 6); Statistics analysis: Student’s t-test analysis; *p<0.05.
Table 14 shows the quantification of H3K9me2 a main repressive mark modulated by G9a (Chen X, Skutt-Kakaria K, Davison J, Ou YL, Choi E, Malik P, Loeb K, Wood B, Georges G, Torok-Storb B, Paddison PJ. G9a/GLP-dependent histone H3K9me2 patterning during human hematopoietic stem cell lineage commitment. Genes Dev. 2012 Nov 15;26(22):2499-511. doi: 10.1101/gad.200329.112. Epub 2012 Oct 26. PMID: 23105005; PMCID: PMC3505820.). Interestingly, we found a reduction of the repressive histone mark H3K9me2 in the hippocampus of SAMP8 treated with Compound 11 at 5 mg/Kg in comparison with SAMP8 Control.
Dendritic length, spine density and Golgi staining protocol
Mice were sacrificed by cervical dislocation and brains were removed from the skull (n=5 whole brain per experimental group). Then, the Golgi staining protocol was developed using the FD Rapid GolgiStain kit according to the manufacturer’s instructions (FD NeuroTechnologies, incs, #PK401). For dendritic branching analysis, images of neurons were collected at 20x magnification in an Olympus BX61 microscope coupled to an Olympus DP70 camera. Measurement of neurite length and complexity was performed using NeuronJ macros and Advanced Sholl Analysis. The number of intersections (branch points) within concentric circles of 10 pm radius was measured and compared between groups. Images for analyzing the spine density were acquired using brightfield microscopy with a 50x oil-objective. All neurites analyzed were around 18 pm and they were at a maximum distance of 150 pm from the soma (Zhang JW, Tabassum S, Jiang JX, Long C. Optimized Golgi-Cox Staining Validated in the Hippocampus of Spared Nerve Injury Mouse Model. Front Neuroanat. 2020 Nov 9;14:585513. doi: 10.3389/fnana.2020.585513. PMID: 33240049; PMCID: PMC7680754.).
Table 15. Quantification of total dendritic length of intersections and spine density in SAMP8 Control and SAMP8 mice treated with Compound 11. (Values presented are mean ± SEM; (SAMP8 Control n = 6, and SAMP8 Compound 11 (5mg/Kg) n = 6); Statistics analysis: Student’s t-test analysis; **p<0.01.
Clauses
For reasons of completeness, various aspects of the invention are set out in the following numbered clauses:
Clause 1. A compound of formula (I), or a salt, solvate or stereoisomer thereof: for use in therapy, wherein:
Ri is CRxR’xRy,
A represents (CH2)m, wherein m is 0 or 1 ,
Z represents (CH2)n, wherein n is 2 or 3,
R2 is selected from the group consisting of: a) an aromatic 5 or 6-membered ring system, wherein the members are selected from the group consisting of: CRzi, S, N, NH, and O; and b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRZ2, S, N, NH, and O;
Rx and R’x are the same or different, and are independently selected from -H, (C1- Cs)alkyl, (Ci-Cs)haloalkyl;
Ry is selected from -H, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, or an aromatic known ring system, which is selected from the group consisting of:
(a) an aromatic 6-membered ring system, wherein the members are selected from the group consisting of: CRZ3, and N;
(b) an aromatic 5-membered ring system, wherein the members are selected from the group consisting of: CRZ4, S, N, NH, and O;
(c) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings having 5 or 6 members selected from the group consisting of: CRzs, N and NH;
Rzi to RZ3 and Rzs are independently selected from the group consisting of: H; OH; CN; halogen; (Ci-Cw)alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more SU3; -0-(Ci-Cio)alkyl; -O-(Ci-Cw)haloalkyl; SF5; S(O)2Rs; NR4R5; CONReR?; and COORs;
RZ4 is selected from the group consisting of: H; OH; CN; halogen; (Ci-Cw)haloalkyl; -O- (Ci-Cw)alkyl; -0-(Ci-Cio)haloalkyl; SF5; S(O)2R’3; NR^R’s; CONR’eR’?; and COOR’s; Rs and R’3 are independently selected from (Ci-Cw)alkyl; (C3-C8)cycloalkyl; aryl; or heteroaryl; 4, Rs, Re, R7, R’4, R’s, R’e and R’7, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl;
Rs and R’s are independently selected from the group consisting of: selected from the group consisting of -H, (Ci-C )alkyl; (C3-Cs)cycloalkyl; and (Ci-Cw)haloalkyl;
“aryl” means an aromatic ring system comprising 6 CRc members, being Rc selected from H, halogen, CN, nitro, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, -O-(Ci-Cs)alkyl, or -O-(Ci- Cs)haloalkyl;
“heteroaryl” means an aromatic ring system comprising 5 or 6 members selected from the group consisting of: CRd, O, N, NH, and S; being Rd selected from H, halogen, CN, nitro, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, -O-(Ci-Cs)alkyl, -O-(Ci-Cs)haloalkyl;
Sui to Su3 are independently selected from the group consisting of: halogen, CN, nitro, (Ci-Ce)haloalkyl, (Ci-Ce)alkyl, (Ci-Ce)hydroxyalkyl, (Ci-Ce)alkoxy, (Ci-Ce)alkylsulfinyl, (Ci-Ce)alkylsulfonyl, (Ci-Ce)alkylcarbonyl, (Ci-Ce)alkoxycarbonyl, carbamoyl, /\/-(Ci- C4)alkylcarbamoyl, /V,/V-di-(Ci-C4)alkylcarbamoyl, (Ci-Ce)alkylcarbonyloxy, (C3- C6)cycloalkyl, phenyl, benzyl, phenoxy, benzyloxy, anilino, /V-methylanilino, phenylmercapto, phenylsulfonyl, phenylsulfinyl, sulfamoyl, /V-(Ci-C4)alkylsulfamoyl, and /V,/V-di-(Ci-C4)alkylsulfamoyl.
Clause 2. The compound for use of clause 1 , wherein m is 1.
Clause 3. The compound for use of any one of the preceding clauses, wherein n is 3.
Clause 4. The compound for use of any one of the preceding clauses, wherein Rx and R’x are the same.
Clause 5. The compound for use of clause 4, wherein Rx and R’x are H.
Clause 6. The compound for use of any one of the preceding clauses, wherein Ry is an aromatic 6-membered ring system. Clause 7. The compound for use of clause 6, wherein the aromatic ring consists of 6 members, which are the same or different, and are CRZ3.
Clause 8. The compound for use of any one of the clauses 6-7, wherein Ry represents phenyl.
Clause 9. The compound for use of any one of the preceding clauses, wherein R2 is an aromatic 5 or 6-membered ring system.
Clause 10. The compound for use of clause 9, wherein R2 is an aromatic 6-membered ring system.
Clause 11. The compound for use of any one of the preceding clauses, wherein Rzi is selected from H; OH; halogen; (Ci-C )alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (C1- Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more SU3; -0-(Ci-Cio)alkyl; -O-(Ci- Cw)haloalkyl; or NR4R5.
Clause 12. The compound for use of clause 11, wherein Rzi is selected from H; OH; halogen; (Ci-Cw)alkyl; -0-(Ci-Cio)alkyl; or NR4R5.
Clause 13. The compound for use of any one of the preceding clauses, which is selected from the group consisting of: 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1,4-oxazepane, 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1-yl]methyl}-1,4-oxazepane, 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1,4-oxazepane, 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1,4-oxazepane,
3-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane, 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline;
3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile,
2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline,
4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol, methyl 3-{ 1 -[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate,
4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1-yl)methyl}-1 ,4-oxazepane,
2-{[4-(1/7-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
3-{1-[(4-(4-chlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
3-{1-[(4-(4-methylbenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine,
4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine,
4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine,
4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine,
2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-methylphenol,
5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-fluorophenol,
3-{1-[(4-(4-methoxybenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
3-{1-[(4-(3,4-dichlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
4-methyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane, 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol,
4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, and any salt, solvate or stereoisomer thereof.
Clause 14. The compound as defined in any one of the preceding clauses 1-13 for use in preventing or treating an EHMT2-mediated disorder in a subject in need thereof.
Clause 15. The compound for use of clause 14, wherein the disorder is associated with overexpression of EHMT2.
Clause 16. The compound for use of any one of the preceding clauses 14-15, which is administered in combination with one or more further therapeutic agents for preventing or treating a disorder associated with overexpression of EHMT2.
Clause 17. The compound for use of any one of the preceding clauses 14-16 for use in the treatment and/or prevention of a disease selected from the group consisting of Alzheimer’s disease, amyotrophic lateral sclerosis, age-related tau astrogliopathy, aortic amyloidosis, argyrophilic grain disease, British familial dementia, cardiac amyloidosis, cerebral amyloid angiopathy, chronic traumatic encephalopathy, corneal dystrophies, corticobasal degeneration, Creutzfeldt-Jakob disease, Danish familial dementia, Down syndrome, familial amyloidosis, familial corneal amyloidosis, fatal insomnia, frontotemporal dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, hereditary cerebral hemorrhage with amyloidosis, Huntington’s disease, inflammation-associated amyloidosis, kuru, Lewy bodies dementia, Mediterranean fever, Niemann-Pick disease type C, Parkinson’s disease, Pick’s disease, primary age-related tauopathy, progressive subcortical gliosis, progressive supranuclear palsy systemic amyloidosis, including transthyretin-associated amyloidosis and light-chain amyloidosis, subacute sclerosing panencephalitis and tuberous sclerosis.
Clause 18. The compound for use of any one of the preceding clauses 14-17 for use in preventing or treating a blood disorder (sickle cell anemia or b-thalassemia or hematological cancer') in a subject in need thereof.
Clause 19. The compound for use of any one of the preceding clauses 14-18 for use in preventing or treating a cancer (lymphoma, leukemia, melanoma, breast cancer, ovarian cancer, hepatocellular carcinoma, prostate carcinoma, lung cancer, brain cancer, or hematological cancer) in a subject in need thereof.
Clause 20. The compound for use of any one of the preceding clauses 14-19, for use in preventing or treating an immune-mediated disease.
Clause 21. The compound for use of any one of the preceding clauses 14-20, for use in reducing substance (alcohol) consumption by a subject.
Clause 22. The compound for use of any one of the preceding clauses 16-18 for use in preventing or treating an imprinting disorder (Prader-Willi syndrome, transient neonatal diabetes mellitus, Silver-Russell syndrome, Albright hereditary osteodystrophy, pseudohypoparathyroidism, Birk-Barel mental retardation, Beckwith-Wiedemann syndrome, Temple syndrome, Kagami-Ogata syndrome, Angelman syndrome, precocious puberty, Schaaf-Yang syndrome, sporadic pseudohypoparathyroidism lb, or maternal uniparental disomy of chromosome 20 syndrome).
Clause 23. A pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in any one of the clauses 1-13, and one or more pharmaceutically acceptable excipients or carriers.
Clause 24. A compound of formula (Ibis), a salt, solvate or stereoisomer thereof: wherein Ri, A, and Z are as defined in any one of the preceding clauses 1-13, and
R2bis is selected from the group consisting of: a) an aromatic 6-membered ring system, wherein the members are selected from the group consisting of: CRzi, and N; and b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRZ2, S, N, NH, and O; CRzi and CRZ2 are as defined in any of the preceding clauses; provided that R2bis is other than 3-hydroxyphenyl.
Clause 25. The compound of clause 24, wherein R2bis is an aromatic 6-membered ring system.
Clause 26. The compound of any one of the clauses 24-25, wherein R2bis is an aromatic 6-membered ring system, wherein all the members are the same or different and represent CRzi, wherein Rzi is as defined in any one of the preceding clauses.
Clause 27. The compound of any one of the clauses 24-26, wherein m is 1 .
Clause 28. The compound of any one of the clauses 24-27, wherein n is 3.
Clause 29. The compound of any one of the clauses 24-28, wherein Rx and R’x are the same.
Clause 30. The compound of clause 29, wherein Rx and R’x are H.
Clause 31. The compound of any one of the clauses 24-30, wherein Ry is an aromatic 6-membered ring system.
Clause 32. The compound of clause 31 , wherein the aromatic ring consists of 6 members, which are the same or different, and are CRZ3.
Clause 33. The compound of any one of the clauses 31-32, wherein CRZ3 represents - CH.
Clause 34. The compound of any one of the clauses 24-33, wherein Rzi is selected from H; OH; halogen; (Ci-C )alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more Sus; -0-(Ci-Cio)alkyl; -O-(Ci-Cw)haloalkyl; or NR4R5.
Clause 35. The compound of clause 34, wherein Rzi is selected from H; OH; halogen; (Ci-Cw)alkyl; -0-(Ci-Cio)alkyl; or NR4R5. Clause 36. The compound of any one of the clauses 24-35, which is selected from:
1. 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1 ,4-oxazepane,
2. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
3. 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4. 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
5. 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
6. 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
7. 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
8. 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
9. 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
10. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
11 . 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
12. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
13. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
14. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline;
15. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile,
16. 2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
17. 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
18. 3-{ 1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline,
19. 4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
20. 4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
21. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
22. 4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
23. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
24. methyl 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate,
25. 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1-yl)methyl}-1 ,4-oxazepane,
26. 2-{[4-(1/7-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
27. 4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
28. 2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
29. 3-{1-[(4-(4-chlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
30. 3-{1-[(4-(4-methylbenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
31. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine,
32. 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
33. 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine,
34. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine, 35. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine.
36. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
37. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-methylphenol,
38. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-fluorophenol,
39. 3-{1-[(4-(4-methoxybenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
40. 3-{1-[(4-(3,4-dichlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
41. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
42. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
43. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
44. 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
45. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
46. 4-methyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
47. 2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
48. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol,
49. 4-{1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, or any salt, solvate or stereoisomer thereof.
Clause 37. A process for preparing the compounds of formula (I) as defined in any of the preceding clauses, which comprises the reaction of a compound of formula (II) with one of formula (III): wherein “X”, represents a leaving group such as halogen or sulfonate (e.g. such as mesylate, tosylate, triflate or the like), particularly X represents Cl; and A, Z, Ri and R2 are as defined in the first aspect of the invention.

Claims

1. A compound of formula (I), or a salt, solvate or stereoisomer thereof: for use in therapy, wherein:
Ri is CRxR’xRy,
A represents (CH2)m, wherein m is 0 or 1 ,
Z represents (CH2)n, wherein n is 2 or 3,
R2 is selected from the group consisting of: a) an aromatic 5 or 6-membered ring system, wherein the members are selected from the group consisting of: CRzi, S, N, NH, and O; and b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRZ2, S, N, NH, and O;
Rx and R’x are the same or different, and are independently selected from -H, (C1- Cs)alkyl, (Ci-Cs)haloalkyl;
Ry is selected from -H, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, or an aromatic known ring system, which is selected from the group consisting of: a) an aromatic 6-membered ring system, wherein the members are selected from the group consisting of: CRZ3, and N; b) an aromatic 5-membered ring system, wherein the members are selected from the group consisting of: CRZ4, S, N, NH, and O; c) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings having 5 or 6 members selected from the group consisting of: CRzs, N and NH;
Rzi to RZ3 and Rzs are independently selected from the group consisting of: H; OH; CN; halogen; (Ci-Cw)alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2-C )alkynyl substituted with one or more SU3; -0-(Ci-Cio)alkyl; -O-(Ci-Cw)haloalkyl; SF5; S(O)2Rs; NR4R5; CONReR?; and COORs; RZ4 is selected from the group consisting of: H; OH; CN; halogen; (Ci-Cw)haloalkyl; -O- (Ci-Cw)alkyl; -O-(Ci-Cw)haloalkyl; SF5; S(O)2R’3; NR^R’s; CONR’eR’?; and COOR’s;
R3 and R’3 are independently selected from (Ci-Cw)alkyl; (C3-C8)cycloalkyl; aryl; or heteroaryl;
R4, Rs, Re, R7, R’4, R’s, R’e and R’7, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl;
Rs and R’s are independently selected from the group consisting of: selected from the group consisting of -H, (Ci-Cw)alkyl; (C3-Cs)cycloalkyl; and (Ci-Cw)haloalkyl;
“aryl” means an aromatic ring system comprising 6 CRc members, being Rc selected from H, halogen, CN, nitro, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, -O-(Ci-Cs)alkyl, or -O-(Ci- Cs)haloalkyl;
“heteroaryl” means an aromatic ring system comprising 5 or 6 members selected from the group consisting of: CRd, O, N, NH, and S; being Rd selected from H, halogen, CN, nitro, (Ci-Cs)alkyl, (Ci-Cs)haloalkyl, -O-(Ci-Cs)alkyl, -O-(Ci-Cs)haloalkyl;
Sui to Su3 are independently selected from the group consisting of: halogen, CN, nitro, (Ci-Ce)haloalkyl, (Ci-Ce)alkyl, (Ci-Ce)hydroxyalkyl, (Ci-Ce)alkoxy, (Ci-Ce)alkylsulfinyl, (Ci-Ce)alkylsulfonyl, (Ci-Ce)alkylcarbonyl, (Ci-Ce)alkoxycarbonyl, carbamoyl, /\/-(Ci- C4)alkylcarbamoyl, /V,/V-di-(Ci-C4)alkylcarbamoyl, (Ci-Ce)alkylcarbonyloxy, (C3- C6)cycloalkyl, phenyl, benzyl, phenoxy, benzyloxy, anilino, /V-methylanilino, phenylmercapto, phenylsulfonyl, phenylsulfinyl, sulfamoyl, /V-(Ci-C4)alkylsulfamoyl, and /V,/V-di-(Ci-C4)alkylsulfamoyl.
2. The compound for use of claim 1 , wherein m is 1 and n is 3.
3. The compound for use of any one of the preceding claims, wherein Rx and R’x are the same, particularly Rx and R’x are H.
4. The compound for use of any one of the preceding claims, wherein Ry is an aromatic 6-membered ring system, particularly Ry represents phenyl.
5. The compound for use of any one of the preceding claims, wherein R2 is an aromatic
5 or 6-membered ring system, particularly R2 is an aromatic 6-membered ring system. 6. The compound for use of any one of the preceding claims, wherein Rzi is selected from H; OH; halogen; (Ci-Cio)alkyl; (C2-C )alkenyl; (C2-C )alkynyl; (Ci-Cw)alkyl substituted with one or more Sui; (C2-C )alkenyl substituted with one or more SU2; (C2- Cw)alkynyl substituted with one or more SU3; -0-(Ci-Cio)alkyl; -O-(Ci-Cw)haloalkyl; or NR4R5; particularly Rzi is selected from H, OH, halogen, (Ci-Cw)alkyl, -0-(Ci-Cio)alkyl, or NR4R5.
7. The compound for use of any one of the preceding claims, which is selected from the group consisting of:
1. 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1 ,4-oxazepane,
2. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
3. 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4. 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
5. 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
6. 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
7. 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
8. 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
9. 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
10. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
11. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
12. 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
13. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
14. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
15. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline;
16. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile,
17. 2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
18. 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
19. 3-{ 1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline,
20. 4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
21 . 4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1 -yl]methyl}- 1 ,4-oxazepane,
22. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
23. 4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
24. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
25. methyl 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate,
26. 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1 -yl)methyl}-1 ,4-oxazepane,
27. 2-{[4-(1/7-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1,4-oxazepane,
28. 4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
29. 2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
30. 3-{1-[(4-(4-chlorobenzyl)-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
31. 3-{1-[(4-(4-methylbenzyl)-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
32. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine,
33. 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
34. 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine,
35. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine,
36. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine.
37. 2-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
38. 5-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-methylphenol,
39. 5-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-fluorophenol,
40. 3-{1-[(4-(4-methoxybenzyl)-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
41. 3-{1-[(4-(3,4-dichlorobenzyl)-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
42. 3-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
43. 3-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
44. 4-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
45. 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1,4-oxazepane,
46. 4-{1-[(4-methyl-1,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
47. 4-methyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
48. 2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1,4-oxazepane,
49. 3-{1-[(4-benzyl-1,4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol,
50. 4-{1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, and any salt, solvate or stereoisomer thereof.
8. The compound as defined in any one of the preceding claims 1-7 for use in preventing or treating an G9a-mediated disorder in a subject in need thereof, particularly by inhibiting G9a.
9. The compound for use of claim 8 for use in preventing or treating a blood disorder, cancer, an immune-mediated disease, an imprinting disorder, or in reducing substance (alcohol) consumption by a subject.
10. The compound for use of any one of the preceding claims 8-9 for use in the treatment and/or prevention of a disease selected from the group consisting of Alzheimer’s disease, amyotrophic lateral sclerosis, age-related tau astrogliopathy, aortic amyloidosis, argyrophilic grain disease, British familial dementia, cardiac amyloidosis, cerebral amyloid angiopathy, chronic traumatic encephalopathy, corneal dystrophies, corticobasal degeneration, Creutzfeldt-Jakob disease, Danish familial dementia, Down syndrome, familial amyloidosis, familial corneal amyloidosis, fatal insomnia, frontotemporal dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, hereditary cerebral hemorrhage with amyloidosis, Huntington’s disease, inflammation-associated amyloidosis, kuru, Lewy bodies dementia, Mediterranean fever, Niemann-Pick disease type C, Parkinson’s disease, Pick’s disease, primary age-related tauopathy, progressive subcortical gliosis, progressive supranuclear palsy systemic amyloidosis, including transthyretin-associated amyloidosis and light-chain amyloidosis, subacute sclerosing panencephalitis and tuberous sclerosis.
11. A pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in any one of the claims 1-10, and one or more pharmaceutically acceptable excipients or carriers.
12. A compound of formula (Ibis), a salt, solvate or stereoisomer thereof: wherein Ri, A, and Z are as defined in any one of the preceding claims 1-11 , and
R2bis is selected from the group consisting of: a) an aromatic 6-membered ring system, wherein the members are selected from the group consisting of: CRzi, and N; and b) an aromatic fused ring system consisting of two rings, wherein one or both rings are aromatic rings, and each one of the aromatic rings have 5 or 6 members selected from the group consisting of CRZ2, S, N, NH, and 0;
CRzi and CRZ2 are as defined in any of the preceding claims; provided that the compound of formula (Ibis) is other than 3-{1-[(4-benzyl-1 ,4-oxazepan- 2-yl)methyl]piperidin-4-yl}phenol. 13. The compound of claim 12, wherein R2bis is an aromatic 6-membered ring system, wherein all the members are the same or different and represent CRzi, wherein Rzi is as defined in any of the preceding claims.
14. The compound of any one of the claims 12-13, which is selected from:
1. 4-benzyl-2-[(4-phenylpiperidin-1-yl)methyl]-1 ,4-oxazepane,
2. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
3. 4-benzyl-2-{[4-(4-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
4. 4-benzyl-2-{[4-(4-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
5. 4-benzyl-2-{[4-(3-bromophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
6. 4-benzyl-2-{[4-(3,5-dichlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
7. 4-benzyl-2-{[4-(2-fluorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
8. 4-benzyl-2-{[4-(p-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
9. 4-benzyl-2-{[4-(m-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
10. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
11 . 4-benzyl-2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
12. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
13. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
14. 4-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline;
15. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzonitrile,
16. 2-{[4-(1/7-pyrazol-1-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
17. 3-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
18. 3-{ 1 -[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline,
19. 4-benzyl-2-{[4-(3-methylsulfonylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
20. 4-benzyl-2-{[4-(pyridin-4-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
21. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
22. 4-benzyl-2-{[4-(3-chlorophenyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
23. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
24. methyl 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}benzoate,
25. 4-benzyl-2-{4-(pyrimidin-2-yl)piperidin-1-yl)methyl}-1 ,4-oxazepane,
26. 2-{[4-(1/7-benzo[d]imidazol-2-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
27. 4-benzyl-2-{[4-(pyridin-3-yl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
28. 2-{[4-(1/7-indol-3-yl)piperidin-1-yl]methyl}-4-benzyl-1 ,4-oxazepane,
29. 3-{1-[(4-(4-chlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
30. 3-{1-[(4-(4-methylbenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
31. 4-benzyl-2-{[4-(4-chlorophenyl)piperidin-1-yl]methyl}morpholine,
32. 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}phenol,
33. 4-benzyl-2-{[4-phenylpiperidin-1-yl]methyl}morpholine,
34. 4-benzyl-2-{[4-(3-trifluoromethylphenyl)piperidin-1-yl]methyl}morpholine,
35. 4-benzyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}morpholine.
36. 2-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
37. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-methylphenol,
38. 5-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}-2-fluorophenol,
39. 3-{1-[(4-(4-methoxybenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
40. 3-{1-[(4-(3,4-dichlorobenzyl)-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
41. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
42. 3-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
43. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}phenol,
44. 2-{[4-(4-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
45. 4-{1-[(4-methyl-1 ,4-oxazepan-2-yl)methyl]piperidin-4-yl}aniline,
46. 4-methyl-2-{[4-(o-tolyl)piperidin-1-yl]methyl}-1 ,4-oxazepane,
47. 2-{[4-(3-methoxyphenyl)piperidin-1-yl]methyl}-4-methyl-1 ,4-oxazepane,
48. 3-{1-[(4-benzyl-1 ,4-oxazepan-2-yl)methyl]pyrrolidin-3-yl}phenol,
49. 4-{1-[(4-benzylmorpholin-2-yl)methyl]piperidin-4-yl}aniline, or any salt, solvate or stereoisomer thereof.
EP24736766.7A 2023-06-22 2024-06-24 Morpholine and 1,4-oxazepane compounds and its use in therapy Pending EP4731223A1 (en)

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